Synchronization of ovulation and fertility in weaned sows treated with intravaginal triptorelin is influenced by timing of administration and follicle size

A 100 μg dose of triptorelin was tested for synchronizing ovulation in sows. In Experiment 1, conducted in April through June, sows (n = 125) were assigned to Control (untreated), TG-96 (Triptorelin Gel (TG) given intravaginally at 96 h post-weaning), or TG-E (given intravaginally at estrus). To optimize AI timing, sows were inseminated at 2 and 26 h after estrus for Control and TG-E and at 8 and 32 h following TG-96. Ovulation by 48 h post-treatment tended to be affected by treatment (P = 0.08) and more (P < 0.05) TG-96 sows ovulated (57.9%) compared to Controls (34.2%), but TG-E (45.1%) did not differ (P > 0.10). Duration of estrus was reduced (P < 0.005) in TG-96 (51 h) and TG-E (58 h) compared to Controls (65 h). There was no treatment effect on farrowing rate (71%) or total born (10.4). Average follicle size <6.5 mm at 96 h after weaning was associated with reduced (P < 0.01) estrus, ovulation and farrowing rate. Experiment 2 was conducted in August through September using 503 weaned sows. The TG-96 treatment reduced duration of estrus (P = 0.03), but treatment did not affect estrus expression, farrowing rate or total pigs born. In conclusion, use of a 100 μg dose of triptorelin intravaginally at 96 h or at estrus advanced ovulation and when used with timed insemination, resulted in similar farrowing rates and litter sizes comparable to sows mated based on estrus. However, ovulation induction and timed AI success may benefit from an approach that ensures sows have adequate follicle development at time of treatment.     

Induction and synchronization of ovulations of nulliparous and multiparous sows with an injection of gonadotropin-releasing hormone agonist (Receptal)

The objective of this study was to determine if administration of a set dose (10 μg) of a gonadotropin-releasing hormone agonist, buserelin (Receptal; Rc), at set times after altrenogest (Regumate; RU) treatment or after weaning was able to induce and synchronize ovulation in female swine (gilts and sows). The pubertal (n = 187) gilts were allocated to four groups, all synchronized with RU. Group 1 (RU) was inseminated twice at detected estrus, Group 2 (RU+Rc120) and Group 4 (RU+Rc104) received 10 μg Rc at 120 or 104 h after the end of RU treatment, respectively, and Group 3 (RU+eCG+Rc104) was treated with 800 IU equine chorionic gonadotropin (eCG) at 24 h and Rc 104 h after the end of RU treatment, respectively. Gilts were inseminated twice at predetermined times, namely 144 and 168 h (Group 2), 128 and 144 h (Group 3), and 144 and 152 h (Group 4) after the end of RU treatment, respectively. Pregnant gilts were slaughtered at 30 d. Administration of Rc 104 h after the end of RU feeding synchronized ovulation over a 24-h time window in 97.9% and 100% of the gilts of Groups 3 and 4, respectively, whereas Rc administration at 120 h (Group 2) only successfully synchronized 88.9% of the gilts over 24 h. Ovulation rates of gilts of Groups 2 and 4 were similar to that of the control group. Pregnancy rates were numerically higher in Groups 2 and 3 (92% and 96%, respectively) compared with those of Groups 1 and 4 (84% and 81%, respectively). Combination of eCG with Rc administration at 104 h (Group 3) increased ovulation rate (+4 CL) but decreased embryo survival to 62% at Day 30. The weaned sow experiment involved 61 sows of a range of parities (2.7 ± 0.9), allocated to two control groups (Control 104 group and Control 94 group) and two treated groups (Rc104 group and Rc94 group), which received 10 μg Rc at 104 and 94 h after weaning, respectively. The females were inseminated at detected estrus. All pregnant sows farrowed. After treatment with Rc 94 h after weaning, 100% of sows ovulated over a 24-h time window versus only 68.7% of controls. Farrowing rate and litter size of the sows treated with Rc at that time were unaffected compared with that of control sows. In contrast, Rc administration at 104 h after weaning may have been too late; only 66.7% of the treated sows ovulated during a 24-h period. This proportion was numerically lower but not significantly different than that for control sows. Farrowing rate and litter size of treated sows were not significantly different than that of controls. Administration of Rc at the dose and times selected in this study tightened synchrony of ovulation in gilts and in sows after weaning. It remains to be established if such a synchrony is suitable to obtain good fertility after a single artificial insemination at a predetermined time.     

Administration of sulpiride or domperidone for advancing the first ovulation in deep anestrous mares

Since results with using sulpiride and domperidone are conflicting and since both have not been tested at the same time, the aim of this study was to compare the efficacy of these substances for the induction of ovulation in deep anestrous mares in the same experimental conditions and to determine their fertility after artificial insemination (AI) at the induced estrus. Twenty-six non-pregnant, non-lactating standardbred anestrous mares were randomly assigned to three groups and treated daily for 25 days (from February 3rd to February 28th) with either sulpiride (1 mg/kg of body weight im SID, n = 10), or domperidone (1 mg/kg po SID, n = 10); 6 animals were used as control. The beginning of the transition period and the first ovulation were hastened in sulpiride (16.4 ± 0.8 days) but not in domperidone (46.0 ± 3.3 days) treated mares (P < 0.05). The diameter of the largest follicle was affected by treatment, time and interaction of treatment-by-day (P < 0.05) and significantly increased in the sulpiride group (P < 0.05). Although a main effect of treatment on plasma LH concentration was not observed (P = 0.06), time and interaction of treatment-by-day were statistically significant (P < 0.05). The interval from the beginning of treatment to first ovulation was shorter (P < 0.05) in the sulpiride group (36.9 ± 2.5 days) than in the domperidone (74.7 ± 3.3 days) and control (81.4 ± 3.1) groups. The establishment of pregnancy was significantly (P < 0.05) hastened in sulpiride (61.0 ± 35.2 days) but not in domperidone (83.0 ± 44.0 days) treated mares. Treated mares not pregnant after the first AI, showed normal estrous cycles with regular interovulatory intervals (P > 0.05). It was concluded that sulpiride is effective in advancing the beginning of transition period and the first ovulation whereas domperidone is successful only in some mares.     

Seasonal fluctuations in the response of Italian Mediterranean buffaloes to synchronization of ovulation and timed artificial insemination

A comprehensive study of the efficiency of synchronization of ovulation and timed artificial insemination (TAI) was undertaken in a large group of Italian Mediterranean buffaloes at a commercial dairy. A total of 2791 synchronization protocols were carried out on 857 animals over 3 years. Of these protocols, 823 (29.5%) did not proceed beyond Day 7 (due to the absence of a vascularized CL) and 620 (22.2%) were discontinued on Day 10 (due to the absence of follicles >1.0 cm and tonic uteri); hence, 1443 (51.7%) protocols did not progress to TAI. Data were analyzed for four periods: P1, transition to spring (from breeding season to low breeding season); P2, low breeding season; P3, transition to fall (low breeding season to breeding season); and P4, breeding season. No differences were found among the four periods in terms of the proportion of protocols that did not result in TAI. Of the 857 buffaloes, 660 (77%) conceived and delivered a calf. The average number of TAI per pregnancy was 2.1 and ranged from 1.9 to 2.3 across years. Logistic regression analysis showed that buffaloes that calved during P3 had a higher odds ratio for pregnancy (1.380; P < 0.05) than buffaloes that calved in other periods. Pregnancy was also influenced by the calving to service period (odds ratio = 0.977; P < 0.01) and the pregnancy per AI (P/AI) at successive TAI (odds ratio = 1.480; P < 0.01). The pregnancy per AI at the first TAI (424/857, 49.5%) was greater (P < 0.01) than in subsequent TAI. The occurrence of late embryonic mortality (between Days 27 and 45 after TAI) was similar among the four periods. These findings indicated that there are distinct seasonal differences in the response of Italian Mediterranean buffaloes to synchronization and TAI.     

The effect of porcine luteinizing hormone in the synchronization of ovulation and corpus luteum development in nonlactating cows

The objective of this study was to determine the effects of different doses of porcine luteinizing hormone (pLH) versus 100 μg gonadotropin-releasing hormone (GnRH) on ovulatory response (during diestrus and proestrus) and corpus luteum (CL) development in nonlactating cows. In Experiment 1, 75 cows received an intravaginal insert containing 1.9 g progesterone (P4) for 10 d to synchronize estrus (Day 0), with prostaglandin F_2α (PGF) at insert removal. On Day 5, all follicles ≥8 mm were ablated, and on Day 12, cows received 8, 12.5, or 25 mg pLH or 100 μg GnRH. Mean (±SEM) plasma P4 concentrations on Day 12 did not differ among treatments (5.6 ± 0.2 ng/mL). Mean plasma LH concentration was greatest (P < 0.01) in cows given 25 mg pLH (4.3 ± 0.4 ng/mL). The ovulatory response to 25 mg pLH (84%) or 100 μg GnRH (72%) was greater (P < 0.05) than that to 8 mg pLH (32%), but not different from that of 12.5 mg pLH (58%). In Experiment 2, 68 cows were given two injections of PGF 10 d apart to synchronize estrus (Day 0). On Day 7, cows received PGF, and, 36 h later, pLH or GnRH (as in Experiment 1). The interval from treatment to ovulation was most variable in cows given 8 mg pLH; only 65% of these cows ovulated during the initial 27 h versus 88% of cows given 25 mg pLH (P < 0.05). Cows given 25 mg pLH or 100 μg GnRH had larger CL area and greater plasma P4 concentrations (P < 0.05) than that of those given 8 mg pLH. In summary, diestrous cows given 25 mg pLH had the greatest plasma luteinizing hormone concentrations, but ovulatory response did not differ from that of those given 100 μg GnRH. Proestrous cows given 25 mg pLH or 100 μg GnRH had greater CL area and P4 concentrations than that of those given 8 mg pLH.     

Comparison of gonadorelin products in lactating dairy cows: Efficacy based on induction of ovulation of an accessory follicle and circulating luteinizing hormone profiles

This study evaluated whether the four gonadorelin products that are commercially available in the United States produce comparable ovulation responses in lactating cows. Dairy cows at 7 d after last gonadotropin-releasing hormone (GnRH) treatment of Ovsynch (Day 7), with a corpus luteum (CL) ≥15 mm and at least one follicle ≥10 mm, were evaluated for response to GnRH treatment. Selected cows were randomized to receive (100 μg; im): (1) Cystorelin (n = 146); (2) Factrel (n = 132); (3) Fertagyl (n = 140); or (4) Ovacyst (n = 140). On Day 14, cows were examined for ovulation by detection of an accessory CL. Circulating luteinizing hormone (LH) concentrations were also evaluated in some cows after treatment with 100 μg (n = 10 per group) or 50 μg (n = 5 per group) GnRH. Statistical analyses were performed with the procedures MIXED and GLIMMIX of the SAS program. Percentage of cows ovulating differed (P < 0.01) among groups, with that for Factrel being lower (55.3%) than that for Cystorelin (76.7%), Fertagyl (73.6%), or Ovacyst (85.0%). There was no effect of batch, parity, or follicle size on ovulation response, but increasing body condition score decreased ovulation response. There was a much greater LH release in cows treated with 100 μg than in those treated with 50 μg, but there were no detectable differences among products in time to LH peak, peak LH concentration, or area under the LH curve and no treatment effects nor treatment by time interactions on circulating LH profile. Thus, ovulation response to Factrel on Day 7 of the cycle was lower than that for other commercial GnRH products, although a definitive mechanism for this difference between products was not demonstrated.     

Variations in the vulvar temperature of sows during proestrus and estrus as determined by infrared thermography and its relation to ovulation

The prediction of ovulation time is one of the most important and yet difficult processes in pig production, and it has a considerable impact on the fertility of the herd and litter size. The objective of this study was to assess the vulvar skin temperature of sows during proestrus and estrus using infrared thermography and to establish a possible relationship between the variations in vulvar temperature and ovulation. The experimental group comprised 36 crossbred Large White × Landrace females, of which 6 were gilts and 30 were multiparous sows. Estrus was detected twice daily and the temperature was obtained every 6 hours from the vulvar area and from two control points in the gluteal area (Gluteal skin temperature [GST]). A third variable, vulvar–gluteal temperature (VGT) was obtained from the difference between the vulvar skin temperature and the GST values. The animals were divided into two subgroups: group A consisting of 11 animals with estrus detected at 6:00 AM, Day 4 postweaning, and group B comprising seven animals with estrus detected at 6:00 AM, Day 5 post-weaning. Both groups showed a similar trend in the VGT. The VGT increased during the proestrus, reaching a peak 24 hours before estrus in group A and 48 hours before estrus in group B. The VGT then decreased markedly reaching the lowest value in groups A and B, respectively, 12 and 6 hours after estrus. Although the time of ovulation was only estimated on the basis of a literature review, the matching between the temporal variations of the VGT values and the predicted time of the peak of estradiol secretion that ultimately leads to the ovulation processes suggests that the VGT values represent a potential predictive marker of the ovulatory events.     

Peripheral administration of metastin induces marked gonadotropin release and ovulation in the rat

Metastin is a novel peptide that has been isolated from the human placenta as the cognate ligand of the G-protein-coupled receptor OT7T175 (or GPR54). However, its physiological functions have not yet been fully investigated. In the present study, we show that subcutaneous administration of metastin increased the plasma levels of gonadotropins (follicle-stimulating hormone and luteinizing hormone) and induced ovulation in prepubertal female rats that had been pretreated with pregnant mare serum gonadotropin to induce follicle maturation. Furthermore, metastin administration drastically increased the plasma levels of gonadotropins in male rats. This action was abolished by pretreatment with a GnRH antagonist, and was accompanied by induction of c-Fos immunoreactivity in GnRH neurons. These results suggest that s.c. administered metastin induces the release of gonadotropin via activation of the hypothalamic GnRH neurons.     

Lack of an effect of prostaglandin injection at estrus onset on the time of ovulation and on reproductive performance in weaned sows

We evaluated the effects of a PGF2alpha analogue on time of ovulation and reproductive performance in multiparous Camborough sows (n=47). At onset of first post-weaning estrus, sows received either an intravulval injection of 3.75 mg of prostaglandin analogue (PGF) or, served as a non-injected control (CON). Beginning 24 h after the onset of estrus, transcutaneous ultrasonography was carried out every six h to determine time of ovulation. At 36, 54, and 72 h after the onset of estrus, blood samples were taken for progesterone analysis. Weaning-to-estrus (WEI), duration of estrus, ovulation rate and number of live embryos at d 28 of gestation were recorded. Treatment had no effect (P > 0.05) on any parameters measured. Duration of estrus classified as less or greater than the overall mean also had no effect (P > 0.05) on any of the parameters measured. Results indicate that treatment did not advance ovulation nor did it improve reproductive performance in sows. Overall, a negative correlation of WEI with the ovulation rate (P = 0.0005, r = -0.54) was established.     

Endocrine changes,timing of ovulation,ovarian follicular growth and efficacy of a novel protocol (Estradoublesynch) for synchronization of ovulation and timed artificial insemination in Murrah buffaloes (Bubalus bubalis)

Experiments were conducted to investigate (1) ovarian follicular growth, timing of ovulation and associated endocrine changes (progesterone, estrogen, and LH) in cycling, and (2) efficacy in terms of pregnancy rate in cycling and anestrus Murrah buffaloes subjected to the Estradoublesynch protocol (prostaglandin F2α [PGF_2α] 0, GnRH 2, PGF_2α 9, estradiol benzoate, EB 10). Twelve cycling buffaloes were subjected to the Estradoublesynch protocol and observed for ovulation, follicle size, and endocrine changes after EB treatment. Ovulation occurred in 12 of 12 buffaloes (100%) at 48.5 ± 1.6 hours (range, 38.0–56.0) after EB treatment. Plasma LH, total estrogen, and progesterone concentrations were determined in intensive blood samples collected after EB treatment. Peak LH concentration of 34.2 ± 7.7 ng/mL (range, 17.8–178.5) occurred at 18.3 ± 0.8 hours (range, 14.0–22.0) after EB treatment. Peak total estrogen of 50.8 ± 6.9 pg/mL (range, 32.3–82.7) occurred 5.7 ± 1.0 hours (range, 2.0–14.0) after EB treatment. Follicle size was gradually increased from second PGF_2α injection (9.7 ± 0.3 mm; range, 8.0–12.0) until ovulation was detected (12.9 ± 0.4 mm; range, 11.0–15.0). Fourteen cycling and 11 anestrus buffaloes were subjected to the Estradoublesynch protocol, with timed artificial insemination (TAI) 48 and 60 hours after EB treatment, and 58 cycling buffaloes were inseminated after spontaneous estrus (control group). Pregnancy rates were 62% for TAI of cycling buffaloes, 64% for anestrus buffaloes, and 34.5% for control group. Our observations demonstrated that the Estradoublesynch protocol followed by TAI satisfactory enhanced pregnancy rates in both cycling and anestrus buffaloes.     

Effect of oestrus synchronization methods on oestrus behaviour, timing of ovulation and pregnancy rate during the breeding and low breeding seasons in Nili-Ravi buffaloes

The objective of this study was to determine the effect of oestrous synchronization methods on oestrous behaviour, timing of ovulation and pregnancy rate during the breeding and low breeding seasons in Nili-Ravi buffaloes. In Experiment 1, oestrous behaviour and timing of ovulation were determined from (n=34) oestruses. The mean (+/- S.E.M.) time of ovulation after the onset of standing oestrus was greater (P<0.05) in PGF(2alpha)-induced luteolysis (30.6+/-1.5h) compared to Ovsynch buffaloes (15.0+/-0.8h). In Experiment 2, pregnancy rates were compared between two methods of synchronization (detected oestrus and Ovsynch protocol) during the breeding and low breeding seasons. Pregnancy rates of buffaloes bred at detected oestrus (62.5%) or by the Ovsynch protocol (36.3%) during the breeding season did not differ significantly (P>0.05) from those which were inseminated during the low breeding season (55.5%) and (30.4%), respectively. This study demonstrates clearly that (1) timing of ovulation in Nili-Ravi buffalo is about 30h after the onset of standing oestrus and (2) buffaloes can be successfully synchronized with optimum fertility using either PGF(2alpha) alone (detected oestrus) or using (Ovsynch protocol) during low breeding season, to calve during the period when milk availability is short.     

Infrared technology for estrus detection and as a predictor of time of ovulation in dairy cows in a pasture-based system

The development and application of an algorithm to assess the ability of an infrared thermography (IRT) device to predict cows in estrus and about to ovulate was investigated. Twenty cows were synchronized using a controlled internal drug release and PGF2α. Vulval and muzzle temperatures were measured every 12 hours from controlled internal drug release insertion to 32 hours after PGF2α treatment and then every 4 hours until ovulation occurred or until 128 hours after PGF2α treatment (whichever occurred first). Thermal images obtained with a FLIR T620 series infrared camera were analyzed using ThermaCAM Researcher Professional 2.9 software. Cows were also monitored for behavioral signs of estrus and color changes of an Estrotect applied to the tail head of each cow 36 hours after PGF2α treatment. Algorithms were developed by adjusting body surface temperature of individual animals for ambient temperature and humidity during each observation period, and were expressed as a deviation from the baseline temperature. Of the 20 cows enrolled in this study, 12 (60%) ovulated. An IRT estrus alert was defined using different thresholds (D = 1 °C, 1.25 °C, and 1.5 °C). Sensitivity and specificity to predict estrus depended upon the chosen threshold level. At a threshold D = 1 °C, the highest sensitivity (92%; n = 11) and the lowest specificity (29%) and positive predictive value (64%) were observed. Conversely, D = 1.5 °C resulted in sensitivity of 75%, specificity of 57%, and positive predictive value of 69%. The mean ± standard deviation intervals between onset and the end of IRT estrus alert to ovulation were 30.7 ± 8.2 and 13.3 ± 7.7 hours, respectively. Ovulation occurred 24 to 47 hours after the onset of the IRT estrus alert for eight out of the 11 ovulated cows (73%). Although the sensitivity of the IRT alert was greater than visual observation (67%) and Estrotect activation (67%), the specificity and positive predictive value were lower than these two aids (i.e., the IRT overpredicted the incidence of ovulation). Results presented indicate that IRT shows some potential as an estrus detection aid; however, further studies investigating the potential to improve the specificity and capturing data throughout entire 21-day reproductive cycles would be worthwhile.     

Effect of low and high egg yolk concentrations in the semen extender for goat semen cryopreservation

This research aimed to evaluate two concentrations of egg yolk inclusion rates (20 and 2.5%) in the semen extender of goat semen cryopreserved during two seasons of the year. The study was conducted during a light-induced breeding season (Experiment 1), and during the natural breeding season (Experiment 2), in the southern hemisphere. Four ejaculates from each buck (n = 2) were collected in each experiment. After collection, semen was divided, with each sample being diluted in the semen extender – according to the treatments (T1 – 20% egg yolk or T2 – 2.5% egg yolk, using a glucose–EDTA extender). For T1 treatment in Experiment 2, the semen was also washed before the semen cryopreservation process. The semen samples were frozen, and after thawing evaluated for seminal characteristics i.e. sperm motility, vigor, morphology and membrane integrity. The fertilising capacity of the frozen-thawed semen was evaluated following a single artificial insemination 12 h after the onset of estrus in 50 (Experiment 1) and 60 does (Experiment 2). In Experiments 1 and 2, the mean values for sperm motility and membrane integrity of the frozen-thawed semen did not differ between the T1 and T2 treatments. However, the mean sperm vigor and morphological normal sperm were greater (P < 0.05) in T2 than T1 treatment. The fertility rates recorded did not differ between T1 and T2 treatments in Experiment 1, however, it was greater (P < 0.05) in the T2 than in the T1 treatment, in Experiment 2. According to obtained results, it can be recommended to use a glucose–EDTA extender with a low egg yolk concentration (2.5%) inclusion, for superior fertility results in goats.     

Effects of the Gut microbiota on Amygdalin and its use as an anti-cancer therapy: Substantial review on the key components involved in altering dose efficacy and toxicity

Conventional and Alternative Medicine (CAM) is popularly used due to side-effects and failure of approved methods, for diseases like Epilepsy and Cancer. Amygdalin, a cyanogenic diglycoside is commonly administered for cancer with other CAM therapies like vitamins and seeds of fruits like apricots and bitter almonds, due to its ability to hydrolyse to hydrogen cyanide (HCN), benzaldehyde and glucose. Over the years, several cases of cyanide toxicity on ingestion have been documented. In-vitro and in-vivo studies using various doses and modes of administration, like IV administration studies that showed no HCN formation, point to the role played by the gut microbiota for the commonly seen poisoning on consumption. The anaerobic Bacteriodetes phylum found in the gut has a high β-glucosidase activity needed for amygdalin hydrolysis to HCN. However, there are certain conditions under which these HCN levels rise to cause toxicity. Case studies have shown toxicity on ingestion of variable doses of amygdalin and no HCN side-effects on consumption of high doses. This review shows how factors like probiotic and prebiotic consumption, other CAM therapies, obesity, diet, age and the like, that alter gut consortium, are responsible for the varying conditions under which toxicity occurs and can be further studied to set-up conditions for safe oral doses. It also indicates ways to delay or quickly treat cyanide toxicity due to oral administration and, reviews conflicts on amygdalin's anti-cancer abilities, dose levels, mode of administration and pharmacokinetics that have hindered its official acceptance at a therapeutic level.     

Effect of equine chorionic gonadotropin on the efficiency of superovulation induction for in vivo and in vitro embryo production in the cat

The effects of various dosages of equine chorionic gonadotropin (eCG) on superovulation induction for in vivo and in vitro embryo production were examined in stray cats (Felis catus). Cats (n = 286) were allocated into five treatment groups with 0, 50, 100, 200, or 400 IU eCG, followed by 100 IU human chorionic gonadotropin (hCG). In vivo- and in vitro-produced blastocysts were obtained by artificial insemination (AI) and in vitro fertilization (IVF), somatic cell nucleus transfer (SCNT), or parthenogenetic activation (PA). The percentage of cats that developed mature follicles, the percentage of cats with collected embryos, and the mean number of in vivo blastocysts per cat were higher in the 200 IU treatment group (43.9%, 31.8%, and 1.53, respectively) compared with those of the other groups (P < 0.05). The percentage of follicular developed cats, the percentage of cumulus-expanded oocytes, and the mean number of collected cumulus-oocyte complexes per cat in the 200 IU (56.7%, 67.8%, and 26.2, respectively) and 400 IU (53.3%, 64.2%, and 26.7, respectively) groups were higher than those in the other groups (P < 0.05). Furthermore, the percentage of in vitro-produced blastocyst per cleaved embryos and the average cell number of the blastocysts from IVF (52.7% and 125.8, respectively) was higher than those of the blastocysts from PA (30.1% and 85.2) and higher than those of the blastocysts from SCNT (15.3% and 37.5; P < 0.05). In conclusion, the current study demonstrated that in vivo and in vitro embryo production were affected by the dosage of eCG; the best results were obtained with 200 IU.     

Monitoring ovulation and implantation in the cynomolgus macaque (Macaca fascicularis) through evaluations of urinary estrone conjugates and progesterone metabolites: a technique for the routine evaluation of reproductive parameters

Investigations were undertaken to determine the applicability of recently reported specific radioimmunoassays for urinary estrone conjugates and progesterone metabolites for monitoring ovarian function in the cynomolgus macaque (Macaca fasciularis) and other macaque species. Mean estrone conjugate measurements appear to accurately reflect the preovulatory estrogen peak in both conceptive (n = 5) and nonconceptive (n = 6) cycles, as well as to indicate early pregnancy through increases which are significantly elevated by Day + 15 (p less than 0.049) post estrone conjugates peak. The mean luteal phase levels of these progesterone metabolites are significantly elevated by Day + 14 (p less than 0.012) in conceptive cycles when compared to the mean values for nonconceptive cycles.     

Ovarian follicular development in Holstein cows following synchronisation of oestrus with oestradiol benzoate and an intravaginal progesterone releasing insert for 5-9 days and duration of the oestrous cycle and concentrations of progesterone following ovulation

The aim of this study was to determine if the duration of treatment with an intravaginal progesterone releasing insert (IVP(4)) after treatment with oestradiol benzoate (ODB) at the time of insertion and 24 h after removal would affect selected variables including: size of ovarian follicles at the time of removal of inserts, diameter of ovulatory follicles, plasma concentrations of progesterone following ovulation, and duration of the following oestrous cycle. Characteristics of oestrus at a synchronised and spontaneous oestrus were also monitored. Non-lactating Holstein cows were synchronised with an IVP(4) for 5 (n = 10), 7 (n = 10), 8 (n = 9) or 9 (n = 9) days together with injections of ODB at device insertion (2 mg) and 24 h after removal (1 mg). Ultrasonography showed no significant effect of treatment on the day of emergence of preovulatory follicles relative to the day of removal of inserts (overall mean = -4.22 +/- 0.58; P = 0.15) for cows that ovulated within 120 h insert removal (n = 36). Treatment with ODB and an IVP(4) for 5 days reduced the diameter of preovulatory follicles at the time of removal of inserts and for the following 2 days compared to cows treated for 7-9 days (mean difference 2.56 +/- 1.15 mm; P = 0.033) but did not reduce the diameter of the ovulatory follicle (P = 0.21). Day of emergence relative to removal of inserts was associated with the diameter of the ovulatory follicle (R2 = 0.69; P < 0.001). Concentrations of progesterone and the diameter of the corpus luteum following ovulation were not affected by treatment (P > 0.20), but were affected by the diameter of the ovulatory follicle (P < 0.01). Diameter of the ovulatory follicle did not affect interoestrous and interovulatory intervals (P > 0.40). We conclude that treatment with an IVP(4) for 5 compared to 7-9 days with ODB administered at device insertion, and 24 h after removal reduced the diameter of preovulatory follicles at the time of removal of the insert but did not reduce the diameter of the ovulatory follicle or concentrations of progesterone in plasma. Emergence of preovulatory follicles closer to the time of removal of inserts reduced the diameter of the ovulatory follicle when oestrus was induced with ODB. Ovulation of smaller follicles reduced concentrations of progesterone in plasma following ovulation but did not affect oestrous cycle duration.     

Development of an enzyme‐linked immunosorbent assay (ELISA) for Luteinizing Hormone (LH) in the elephant (Loxodonta africana and Elephas maximus)

A simple, rapid enzyme‐linked immunosorbent assay (ELISA) for the measurement of LH in plasma and serum of elephants (Loxodonta africana and Elephas maximus) has been developed, validated, and used for comparative studies. Purified elephant LH (eleLH) diluted in elephant plasma was used as standards (0.78–50 ng/ml). A monoclonal antibody against the β‐subunit of bovine LH (518B₇) was used as the capture antibody. The second antibody (a polyclonal rabbit anti‐human LH antibody), conjugated to horseradish peroxidase, cleaved a substrate (tetramethyl benzidine), resulting in a color change. The total assay time was approximately 2½ hr, with incubations at room temperature. Sensitivity was found to be 1.56 ng/ml. Cross‐reactivities to elephant FSH and TSH were low: 0.9% and 0.15%, respectively. The accuracy of the assay was demonstrated by comparing the ELISA with a validated eleLH radioimmunoassay (RIA), progesterone data, and ultrasound observations. Blood samples from 18 Asian and African elephant cows were analyzed with the ELISA and RIA, and an additional 11 cows were used to describe endocrine parameters for LH and progesterone using only RIA. No difference was found in LH peak concentrations between the ELISA and RIA. The time from the progesterone decline to the first LH peak, and the time between the two peaks were similar between species. Asian cows had higher LH peaks than African cows. Ultrasound confirmed the time of ovulation occurring with the second LH peak. Three cows were inseminated and confirmed to be pregnant using this ELISA as a timing device. Instrumentation is not always required, as LH peaks approximating 3 ng/ml can be visually observed. In conclusion, this ELISA can be used as a field test to determine time of ovulation for artificial insemination (AI) or natural breeding of both species of elephants, and thus is an important tool for the preservation of captive populations worldwide. Zoo Biol 23:65–78, 2004. © 2004 Wiley‐Liss, Inc.