A method of artificial insemination in captive White-tailed deer (Odocoileus virginianus )

Production of fawns by artificial insemination in captive White-tailed deer (Odocoileus virginianus ) has been accomplished by using frozen-thawed spermatozoa. The purpose of this study was to determine if frozen-thawed semen deposited at the posterior face of the os cervix could produce conception. Five hand-raised female White-tailed deer and one hand-raised male White-tailed deer were used over two breeding seasons 1984-1985 and 1985-1986. The vasectomized buck was ued to detect estrus in the does. The does were inseminated with frozen-thawed semen containing at least 100 million live normal cells with a 60% or higher motility. The artificial insemination catheters used in this study worked well, but due to the small size of the cervix, the catheter could only be passed up to the first cervical ring, the site at which the semen was deposited. Over two breeding seasons, nine does were inseminated with frozen-thawed spermatozoa; each doe was inseminated once each estrous cycle at one of the following times: 0, 6, 12, 18, 24 or 30 h. after detection of estrus. Of the nine does inseminated with frozen-thawed spermatozoa, six conceived and carried to term 11 healthy normal fawns, yielding an overall conception rate of 67%.     

首例大熊猫秋季发情、配种、产仔

本文记录一只因环境、饲料等因素改变而2001年春季未能发情的雌性大熊猫,于秋季9月中旬发情,并与另一只同期发情的公熊猫自然交配,当年冬季12月17日成功产仔的这一世界首例圈养大熊猫秋季发情配种的行为表现和配种经过,并对该大熊猫秋季发情的原因作了分析。     

Estrus synchronization with melengestrol acetate in cyclic ewes. Insemination with fresh or frozen semen during the first or second estrus post treatment

A total of 540 cyclic ewes were randomly allocated to 1 of 6 groups according to synchronization or not with melengestrol acetate (MGA), insemination with fresh or frozen semen, and insemination during the first or second estrus post treatment. The MGA was effective in synchronizing estrus, since the percentage of ewes showing estrus during the first 6 days after treatment was significantly higher (P<0.05) for treated (79.5%) than for nontreated ewes (33.5%); 74% of the treated ewes showed estrus during a 72-h period. Progesterone concentrations in plasma samples obtained at random from 34 treated ewes on Day 8 post estrus revealed that 94% of them ovulated and formed a functional CL. Synchronization was maintained during the second estrus post treatment, since 71.9% of the ewes showed the second estrus during a 72-h period. Treatment with 0.22 mg of MGA/head/d for 14 d had a detrimental effect on fertility when insemination was carried out during the first estrus post treatment. Delaying insemination until the second estrus post treatment caused a less marked reduction in conception rates. Thus, MGA can be a useful alternative for estrus synchronization of a large number of ewes. Artificial insemination can be delayed until the second estrus post treatment, improving fertility without loosing the advantages of estrus synchronization.     

大熊猫超轻初生幼仔的成功哺育

大熊猫初生幼仔体重较轻,仅为正常母体体重的千分之一,平均144.9±40.59 g(张志和和魏辅文,2006).初生体重在70 g以下的幼仔,常被称之为超轻体重幼仔,初生体重超轻、身体瘦弱活动无力,产后无叫声或叫声小是其主要特点.对于体重超轻初生幼仔的育幼,目前有1993年对一例初生体重为61 g的幼仔,在产后两周内采用胃导管喂母初乳、之后采取人工与母兽交替哺育的方法养育存活以及2000年对一只58 g幼仔进行全人工育幼存活7 d的报道(钟顺龙和何光昕,1997,侯蓉等,2000).2006年成都大熊猫繁育研究基地的一只雌性大熊猫产下一对双胞胎,其中一仔初生体重仅51 g,经人工辅助哺育存活,该幼仔是圈养条件下育成的初生体重最轻的大熊猫.现将该例的育幼情况报道于后,以供参考.     

Effect of one spontaneous estrus cycle (after synchronization with PGF2alpha) on reproductive performance in dairy cows

The objective of the study was to analyze the effect of a spontaneous estrus cycle after synchronization of estrus with prostaglandin F2alpha (PGF2alpha) in dairy cows on the degree of synchronization and reproductive performance. We assigned 557 Holstein cows to two treatment groups. In Group 1 estrus was synchronized by two treatments with 25 mg of Dinoprost-Trometamol in 14-day intervals. Cows were treated 27 to 33 days postpartum (dpp) and 41 to 47 dpp, respectively. Cows in Group 2 were treated with 25 mg of Dinoprost-Trometamol three times in 14-day intervals, starting at 34 to 40 dpp. The second and third injections were administered at 48 to 54 dpp and 62 to 68 dpp, respectively. All cows were inseminated on observed estrus after a voluntary waiting period of 65 days post partum. Thus cows in Group 1 were inseminated on spontaneous estrus and cows in Group 2 on induced estrus. Cows not inseminated at 80 days post partum were palpated per rectum and treated according to a predefined protocol. Herd reproductive performance measures did not differ significantly between groups. The proportion of cows with low serum progesterone levels was significantly higher 3 days after synchronization than 24 days after synchronization (97% vs 39%). The first-service conception rate was 34.8% in Group 1 and 30.7% in Group 2 (P > 0.05). Days open were 113.5 in Group 1 and 110.9 in Group 2 (P > 0.05). It is concluded that postponing artificial insemination for one spontaneous estrus cycle after synchronization decreased the degree of synchronization. This procedure, however, had no effect on herd reproductive performance compared to insemination on first observed estrus after synchronization.     

Efficacy of a prostaglandin analogue in reproduction in the cycling mare.

A prostaglandin F analogue caused luteolysis in normal cycling non-lactating mares, and lactating mares (treated after the foal estrus). Effective doses ranged from 1.0 to 4.0mg given as a single subcutaneous injection 8–10 days after ovulation. A dose of 0.5mg was ineffective, hence the dose-response relationship was steep, indicative of a quantal type of response. Mares usually returned to estrus within 2–4 days and ovulated by 7 days after treatment. Mares bred naturally or by artificial insemination at the induced estrus and ovulation were fertile. The compound was without side-effects, and hence should be of value in manipulating the estrous cycle of the mare.     

Estrous cycle synchronization in dairy heifers with the prostaglandin analog alfaprostol (I)

The prostaglandin F(2alpha) analog (PGFA) alfaprostol was used in 277 cyclic dairy heifers for the purpose of estrous cycle synchronization. A dose of 5 mg was used in all the trials during spring 1979 and 1.5 mg 100 kg body weight were used during winter 1979/80. Animals were treated according to 3 schedules: two doses 11 days apart without prior examination (Schedule I), one dose without prior observation and a second dose 11 days later only for those animals that failed to show estrus after the first treatment (Schedule II) and one dose for animals not showing estrus after a 5-day observation period (Schedule III). Estrus synchronization was achieved with peak estrus activity occurring form 32 to 72 h after treatment in 95% of the responding animals. Of the animals treated according to schedules I, II and III, 93%, 100% and 100% showed synchronized estrus activities respectively. Conception rates in all trials, from insemination at observed heat (Schedules I, II and III) or from fixed time insemination at 48 and 72 h after the first or second treatments in schedules III and I, respectively, compared well with that of untreated contemporary controls with a range of 34.2 to 66.7% for animals in these trials, and an overall conception rate of 49.48% for treated and of 48.30% for control animals. This observation, together with the pregnancy rate at 60 days after breeding (78.30% for all treated animals and 73.50% for all untreated controls), indicates that alfaprostol had no adverse effects on fertility.     

The economic effects of an estrus synchronization protocol using prostaglandin in beef heifers

We estimated the effect of estrus synchronization on reproduction, production and economic outcomes in 272 beef heifers randomly allocated to a synchronized Test group or an unsynchronized Control group. The Test group received AI upon estrus detection for 6 days followed by PGF2 treatment of heifers that had not shown estrus by day 6 (PGF/6). In both groups AI was continued for 50 days, followed by a 42-day bull breeding period. Heifers were followed through their second breeding season and until they had weaned their first calves. Synchronization resulted in a reduction in median days to first insemination (8 vs. 11 in the Test and Control groups, respectively, P < 0.01) and median days to calving of calves born to AI (14 vs. 20, P = 0.04). There was no significant difference in pregnancy rate to the AI period (60.0% vs. 51.8%, P = 0.18), final pregnancy rate (82.2% vs. 83.2%, P = 0.87) or pregnancy rate to the subsequent breeding season (96.0% vs. 95.0%, P = 1.00). Although mean calf weaning mass was not significantly different (207.0 kg vs. 201.4 kg, P = 0.32), the total mass of calves weaned in this study was 14,843 kg vs. 13,060 kg and the benefit: cost ratio for synchronization was 2.8. It was therefore concluded that a PGF/6 protocol may affect the total mass of calves weaned by changing days to calving, weaning rate, the ratio of male: female calves born and/or the birth mass of calves.     

Protracted reproductive seasonality in the male giant panda (Ailuropoda melanoleuca) reflected by patterns in androgen profiles, ejaculate characteristics, and selected behaviors

The female giant panda (Ailuropoda melanoleuca) experiences a brief (24-72 h) seasonal estrus, occurring once annually in spring (February-May). Our aim was to determine the existence and temporal profile of reproductive seasonality in the male of this species. The study was facilitated by 3 yr of access to eight giant panda males living in a large breeding center in China. Seasonal periods for the male were defined on the basis of female reproductive activity as prebreeding, breeding (early, peak, late), and nonbreeding seasons. Testes size, fecal androgen excretion, ejaculated sperm density, and frequency of reproductive behaviors (i.e., locomotion, scent marking, vocalizations) increased (P < 0.05) from the prebreeding period (October 1-January 31) to the early breeding season (February 1-March 21). Testes volume and sperm concentration were maximal from March 22 through April 15, a period coinciding with maximal female breeding activity. The occurrence of male reproductive behaviors and fecal androgen concentrations began declining during peak breeding and continued from April 16 through May 31 (late breeding period), returning to nadir throughout the nonbreeding interval (June 1-September 30). Reproductive quiescence throughout the latter period was associated with basal testes size/volume and aspermic ejaculates. Our results reveal that testes morphometry, fecal androgen excretion, seminal quality, and certain behaviors integrated together clearly demonstrate reproductive seasonality in the male giant panda. The coordinated increases in testes size, androgen production, sperm density, and sexual behaviors occur over a protracted interval, likely to prepare for and then accommodate a brief, unpredictable female estrus.     

Estrous synchronization in the gaur (Bos gaurus): Behavior and fertility to artificial insemination after prostaglandin treatment

The gaur (Bos guarus) is an endangered species that is a wild ancestor of domestic cattle. This study was conducted to evaluate 1) the efficacy of prostaglandin F_2α (PG) to synchronize estrus in gaur, 2) behavior of male and female gaur around estrus, and 3) fertility after artificial insemination (AI) at the PG-induced estrus. Six female gaur were utilized, along with a vasectomized gaur bull used to aid in detecting estrus. All females were given two i.m. injections of PG (25 mg/injection) 11 days apart, and monitored for estrus for 120 hr (48 hr via chinball marks, followed by 72 hr continuous observation) after the second PG injection. Three of the six females were in estrus during the 120-hr evaluation period. One female was in estrus prior to 48 hr after the second PG injection, and two females were observed in estrus during the 72-hr continuous observation period. When a female was observed in estrus (standing to be mounted by the vasectomized bull), she was bred by AI at 12 and 24 hr after the onset of estrus. The four females not observed in estrus, including the one marked during the first 48 hr, were bred by AI at 80 and 92 hr after the second PG injection. Of the two gaur females observed in estrus, one female was first mated by the vasectomized bull at 77 hr and the second female was mated at 98 hr after PG. Both females exhibited very short durations of receptivity (less than 4 hr). The second female observed in estrus became pregnant after AI and gave birth to a healthy gaur calf after a 299-day gestation. It appears that female gaur can be synchronized with PG techniques developed for domestic cattle. These data should provide useful information for programs studying and maintaining this endangered species and may have relevance for the cattle industry, since the gaur could provide a source of diverse ancestral genetic material.     

Early-weaned sows: altrenogest therapy, estrus, ovulation, and reproductive performance

Early weaning is a technique used to increase swine health status, and may cause consequences in reproductive performance of sows. An experiment was performed to evaluate these effects in a herd of sows, with weaning at 9 or 10 days post-farrowing, located in west of Minas Gerais, Brazil. Large-White sows (n=102), with three or four previous parturitions were randomly allocated to three treatment groups: T1: artificial insemination (AI) at first post-weaning estrus of the sows; T2: AI at second post-weaning estrus, T3: AI at first estrus, after an administration of a daily individual dose of 20 mg of altrenogest from 5 to 8 days post-weaning. The duration of the first post-weaning estrus did not differ among treatment groups; however, the second estrus of the T2 group was of shorter duration relative to the other treatment groups (P< or =0.035). Ovulation occurred earlier at the second estrus of the T2 group, compared with the T1 and T3 groups (P< or =0.027), being similar to that at the first estrus of T2 group (P=0.177). The relationship of the timing between ovulation and estrus was similar among treatment groups (P> or =0.221). There was no difference in farrowing rate among treatment groups (P> or =0.313). The T2 group produced a mean of 2.5 more piglets per litter (P=0.002). In conclusion, the use of altrenogest did not increase the reproductive performance of early-weaned sows.     

Papanicolaou staining of exfoliated vaginal epithelial cells facilitates the prediction of ovulation in the giant panda

The giant panda is seasonally monoestrus, experiencing a single estrous with spontaneous ovulation in the spring. Therefore, accurate monitoring of the estrous cycle to pinpoint the time of ovulation is critical for the success of timed mating or artificial insemination. Analysis of exfoliated vaginal epithelial cells is a simple technique that rapidly yields information about the estrous status of a panda. Vaginal swabs were obtained during five estrous cycles of two nulliparous females. Cells were stained with the trichrome Papanicolaou and classified as basophils, intermediates or superficials. The color of stained cells, basophilic, acidophilic or keratinized, was recorded as a characteristic independent of the three standard cell types. The day urinary conjugates of estrogen fell from peak levels was considered the day of ovulation. A chromic shift occurred 8-9 days before ovulation when the majority of exfoliated vaginal cells changed from basophilic (blue) to acidophilic (pink) without accompanying nuclear or cytoplasmic changes. A second chromic shift was consistently observed 2 days prior to ovulation when keratinized (orange) cells replaced acidophils as the majority of vaginal cells. Monochrome staining of vaginal cells is sufficient to quantify superficial cells, which is a useful adjunct to behavioral and endocrinological data in determining estrous in the giant panda. However, the timing and duration of superficial cell elevations are substantially different between and within individual females, which limits the accuracy of timing ovulation for artificial insemination. The predictive value of vaginal cytology was greatly enhanced with the trichrome stain and evaluation of cell color.     

GnRH-agonist induction of fertile estrus with either natural mating or artificial insemination, followed by birth of pups in gray wolves (Canis lupus)

Although captive populations of endangered species such as the Mexican gray wolf (Canis lupus baileyi) can benefit from artificial insemination to accomplish genetic exchange, reliable techniques for timing insemination are lacking. We used the generic gray wolf (C. lupus) to test the efficacy of a short-acting GnRH-agonist implant, deslorelin, for inducing estrus. Of five females receiving implants on 17 or 18 January 2003, two mated naturally 10-17 days later, and the others were artificially inseminated using fresh semen, one on day 7 and all three on day 11. Relaxin tests revealed that one artificially inseminated female and both naturally mated females were pregnant on 1 March, and all three gave birth to healthy puppies on 4-6 April. Of the artificially inseminated females, only the one who subsequently conceived and gave birth was judged to be in cytologic estrus at the time of insemination. Two females were treated again with deslorelin on 12 January 2004, followed by collection of fecal samples for hormone analysis. One female, who was housed with a male, copulated on day 17 but did not conceive; the other was not with an adult male. Fecal progestin and estrogen profiles suggested that estrus, but not ovulation, was induced. These results indicated that deslorelin could induce fertile estrus in the gray wolf, although individual response varied. Further investigation is needed to better define and control the interval between implant insertion and ovulation for optimal timing of insemination.     

Litter loss triggers estrus in a nonsocial seasonal breeder

Sexually selected infanticide (SSI) is often presumed to be rare among seasonal breeders, because it would require a near immediate return to estrus after the loss of an entire litter during the mating season. We evaluated changes in reproductive strategies and the reproductive fate of females that experienced litter loss during the mating season in a seasonal breeder with strong evidence for SSI, the brown bear. First, we used a long‐term demographic dataset (1986–2011) to document that a large majority of females (>91%) that lose their entire litter during the mating season in fact do enter estrus, mate, and give birth during the subsequent birthing season. Second, we used high‐resolution movement data (2005–2011) to evaluate how females changed reproductive strategies after losing their entire litter during the mating season. We hypothesized that females would shift from the sedentary lifestyle typical for females with cubs‐of‐the‐year to a roam‐to‐mate behavior typical for receptive females in no more than a few (~3) days after litter loss. We found that females with cubs‐of‐the‐year moved at about 1/3 of the rate and in a less bimodal diurnal pattern than receptive females during the mating season. The probability of litter loss was positively related with movement rate, suggesting that being elusive and sedentary is a strategy to enhance cub survival rather than a relic of cub mobility itself. The movement patterns of receptive females and females after litter loss were indistinguishable within 1–2 days after the litter loss, and we illustrate that SSI can significantly reduce the female interbirth interval (50–85%). Our results suggest that SSI can also be advantageous for males in seasonally breeding mammals. We propose that infanticide as a male reproductive strategy is more prevalent among mammals with reproductive seasonality than observed or reported.     

Split parturition observed in a captive North American brown bear (Ursus arctos)

Reproductive physiology in North American ursids is characterized by mating from spring to early summer, delayed implantation, and birth during hibernation. During spring 2008, a captive adult female brown bear was mated with two adult males. Pregnancy was determined by elevated progesterone concentrations during late fall before hibernation. Two male cubs were born on December 31, 2008, and a third female cub was born 17 days later on January 16. All were successfully raised and all were confirmed to have identical paternity. When normalized to age, cub growth rates did not differ. To our knowledge, this is the first documented case of markedly different birth dates in a single litter of brown bear cubs.     

Fertility following fixed-time AI or insemination at observed estrus in Ovsynch and Heatsynch programs in lactating dairy cows

The objective of this study was to compare the conception rate for fixed-timed artificial insemination (FTAI) and observed heat artificial insemination (HAI) prior to the scheduled FTAI in Ovsynch and Heatsynch synchronization protocols. In Experiment 1, lactating dairy cows (n=535) received two set-up injections of 25mg prostaglandin F(2alpha) (PGF(2alpha)) i.m., 14 days apart starting at 36+/-3 days in milk (DIM). Cows were blocked by parity and were randomly allocated to either Ovsynch or Heatsynch groups. All cows received 100 microg of GnRH i.m. 14 days after the second set-up injection of PGF(2alpha), followed by a third injection of 25mg PGF(2alpha) i.m., 7 days later. In the Ovsynch group, HAI cows (n=29) were bred on standing estrus after the third PGF(2alpha) before the scheduled second GnRH, whereas FTAI cows (n=218) that were not observed in estrus, received a second injection of 100 microg of GnRH i.m., 48 h after the third PGF(2alpha) and received TAI 8 h after the second GnRH. In the Heatsynch group, all cows (n=288) received 0.5 mg of estradiol cypionate (ECP) 24 h after third PGF(2alpha) and HAI cows (n=172) were bred on standing estrus and FTAI cows (n=116) that were not observed in estrus, received TAI 72 h after the third PGF(2alpha). In Experiment 2, repeat breeder cows (n=186) were randomly assigned to either Ovsynch or Heatsynch groups. The FTAI and HAI cows were inseminated similar to Experiment 1. All cows were observed for estrus three times daily. The associations with the conception rate were modeled with logistic regression separately for Experiments 1 and 2. Of all the variables included in the model in Experiment 1, type of AI (HAI versus FTAI, P=0.0003) and parity (primiparous versus multiparous, P=0.05) influenced the first service conception rate. Over-all conception rate and first service conception rate for HAI cows were higher compared to FTAI cows (33.8% versus 21.3%, and 35.3% versus 21.0%; P=0.001). In the Heatsynch group, cows that received HAI had significantly higher over-all conception rate and first service conception rate compared to FTAI (35.2% versus 17.3% and 36.0% versus 15.5%; P=0.0001). The conception rates in repeat breeder cows for HAI and FTAI (30.1% versus 22.3%) were not different (P>0.1). In conclusion, it was recommended to include AI at observed estrus and fixed-time AI for cows not observed in estrus in order to improve the conception rate in synchronization protocols.     

Artificial insemination increases the probability of a male calf in dairy and beef cattle

The objective of this study was to determine if natural mating affected secondary sex ratio. Data consisting of 642,401 calving records from the Irish national database, during the years 2002-2005, were used in the analysis. Factors affecting the logit of the probability of a male calf being born were determined using multiple regression generalised estimating equations with sire of the calf included as a repeated effect. Month of the year at calving, sex of the previous calf born within dam, breed of service sire, parity of dam and type of mating (i.e., natural or artificial insemination) significantly (P<0.05) affected the likelihood of a male calf being born. Male calves were more likely to be born in the warmer months of the year, when the sex of the previous calf born to the same dam was male, in older cows and when the service sire was a beef breed. No significant interaction between the main effects existed. The odds of a male calf being born, following adjustment for confounding effects, varied from 1.04 to 1.08 (P<0.01) across the years of analysis when artificial insemination was used compared to natural mating. This equates to a 1% unit increase in the probability of a male calf being born following artificial insemination.     

Reproductive response of ewes synchronized with different lengths of MGA treatments in intrauterine insemination program

A total of 415 fat tailed ewes were randomly assigned to two groups to assess the effect of duration of melengestrol acetate (MGA) (9 versus 12d) administration on reproductive parameters associated with laparoscopic artificial insemination. At the end of MGA treatment, ewes in each group were subdivided and inseminated with one of two different insemination doses (10×10(7) or 20×10(7) sperm per 0.5 ml insemination dose) of fresh diluted semen. Inseminations were carried out 11-18 h after first detected estrus. Ewes were screened for their return to oestrus from 10 to 21 days post AI and inseminated at their returned oestrus. Pregnancy diagnosis was done from approximately 55 days after insemination in both synchronized and return estrus. For short (9-day) and long (12-day) term MGA treated groups, estrus rates were 62% versus 89% (P<0.0001), respectively. Ewes (n=115) that returned to estrus were inseminated (7-11h after estrus detection) with fresh diluted semen at different doses (20×10(7) or 40×10(7) or 60×10(7) sperm per 0.5 ml insemination dose). Pregnancy rates were 41% and 44% for short term and long term MGA treated ewes, respectively. Pregnancy rate of ewes which returned to oestrus was 53.4%. There was a significant (P<0.05) increase in pregnancy rates (38-52% for 11-16 h; 63% for 17-18 h) when insemination was held at 17-18 h after first detected estrus following MGA treatments. Pregnancy rates were found to be similar in ewes inseminated with 10×10(7) (36%) or 20×10(7) (47%) motile spermatozoa at first AI, and 20×10(7) (44%) or 40×10(7) (59%) or 60×10(7)(48%) at second AI. It was concluded that short term MGA treated ewes were recorded with lower estrus rates but was similar to pregnancy rates with long term MGA treatment. Acceptable pregnancy rates were achieved in MGA induced estrus when insemination is conducted at 17-18 h after estrus onset and with 20×10(7) sperm per insemination dose.     

Successful artificial insemination in the koala (Phascolarctos cinereus) using extended and extended-chilled semen collected by electroejaculation

Artificial insemination in the koala using chilled, electroejaculated semen provides for a marked improvement in the reproductive and genetic management of captive koala colonies in Australia and internationally, and makes available the option of using semen collected from wild populations to expand restricted gene pools. Dilution of koala semen for artificial insemination is complicated because koalas are induced ovulators, and it is thought that ovulating factors are present in the semen, so that semen extension for preservation purposes might be anticipated to result in a failure to induce ovulation. The first two experiments of this study were designed to determine whether artificial insemination using undiluted, extended, and extended-chilled semen collected by electroejaculation was capable of inducing a luteal phase and/or the production of pouch young. In Experiment 1, 1 ml undiluted electroejaculated semen, 2 ml diluted (1:1) semen, and 1 ml diluted (1:1) semen resulted in seven of nine, six of nine, and six of nine koalas showing a luteal phase, respectively; four pouch young were produced in each treatment. A second artificial insemination experiment was conducted in which 2 ml diluted (1:1) semen was administered in three groups of nine koalas. The first group received semen that had been collected and diluted immediately without chilling, the second group was deposited with semen stored chilled for 24 h, and the final group received semen that had been chilled for 72 h. In the first group, five females had a luteal phase, but none became pregnant. In group 2, two of the five females that had a luteal phase gave birth, whereas in group 3, four of the six females that had a luteal phase produced pouch young. In addition, experiment 3 was conducted to determine whether it was possible to produce pouch young by naturally mating koalas that were in the latter stages of their behavioral estrus; this information is important to the logistics of transporting koala semen for artificial insemination by establishing the maximum time frame in which females might be expected to shed a fertile oocyte. Of the 12 females mated on Day 8 of estrus, 6 gave birth, whereas only 3 of the 10 females naturally mated on Day 10 of estrus produced pouch young. The majority of females (21 of 22) in experiment 3 showed evidence of a luteal phase. Together, these experiments have shown that it is possible to use undiluted, extended, or extended-chilled semen to produce koala offspring up to Day 8 of estrus at conception rates similar to those achieved following natural mating. These findings represent a significant advancement in the use of reproductive technology in marsupials and provide the basis for the shipment of koala semen over long distances. The pouch young produced in this study represent the first marsupials born following artificial insemination of extended-chilled semen and bring the total number of koalas produced by artificial insemination to 31.     

Photoperiod manipulation to increase the productivity of black‐footed ferrets (Mustela nigripes) and Siberian polecats (M. eversmanii)

The endangered black‐footed ferret (Mustela nigripes) reproduces naturally in the spring. The Black‐Footed Ferret Recovery Program would benefit from increased productivity in the captive population by manipulating photoperiod and temperature to create three artificial cycles in 1½ years. In the present study the photoperiod was set at 8 hr light:16 hr dark for 1–2 months, then switched to 16 hr light:8 hr dark for 4–5 months. The males' light was switched 1 month before the females' light. Three cycles were performed in 2 years. During the first cycle, 0% of the males bred, 17% of the females came into estrus and were bred through artificial insemination, and 0% of the females whelped. In the second cycle, 100% of the females came into estrus and were bred naturally, and 77% whelped. In the third cycle, all males showed testicular recrudescence but none bred; all females showed signs of estrus, 40% were bred using artificial insemination, and 0% whelped. Siberian polecats (Mustela eversmanii), the black‐footed ferrets' closest living relative, also were put on an artificial photoperiod, coinciding with the black‐footed ferrets' third cycle. All female polecats came into estrus and were bred naturally, and 33% whelped. All males showed testicular recrudescence, and 22% produced sperm and bred. The low rate of success in breeding and whelping suggests that multiple cues may be needed to induce estrus in ferrets and polecats. Zoo Biol 22:1–14, 2003. © 2003 Wiley‐Liss, Inc.