Porcine Luteinizing Hormone Research Articles

Morphologic, Steroidogenic, and Transcriptomic Assessment of the Corpus Luteum in Holstein Cows after Spontaneous or Hormone-Induced Ovulation.

There is evidence that replacing the gonadotropin-releasing hormone (GnRH) with porcine luteinizing hormone (pLH) to synchronize ovulation prior to artificial insemination (AI) increased pregnancy per AI in dairy cows without affecting blood progesterone (P4) concentrations. Whether morphologic, steroidogenic, and transcriptomic differences exist among corpora lutea (CL) formed after ovulation induced by GnRH and pLH is unclear. Our main objective, therefore, was to compare CL characteristics between GnRH- and pLH-induced CL. In 24 non-lactating Holstein cows, ovulations were spontaneous (Spont-Ov) or induced with 100 µg GnRH, 25 mg pLH, or 1 mg estradiol benzoate (EB), with CL excised 12 d after ovulation. In pLH- versus GnRH-treated cows, the duration of elevated LH (above baseline) was prolonged (10 versus 6 h, respectively, p < 0.01), but CL dimensions, pixel intensity of CL images, proportions of steroidogenic and non-steroidogenic luteal cells, and mean plasma LH did not significantly differ. Post-ovulation mean plasma P4 (ng/mL) did not differ among Spont-Ov (3.0) pLH (3.1) or GnRH (3.0) cows but were lower in EB cows (2.0). In vitro P4 concentration was greater in luteal explants of pLH-treated cows than in all other groups (combined means, 16.0 vs. 12.3 µg/mL, p < 0.02). Relative abundance of mRNA for oxytocin receptor (OXTR) was 2-fold higher (p < 0.01) in CL of pLH vs. GnRH cows and highest in Spont-Ov CL. In summary, pLH-treated cows had a longer LH peak, and greatest luteal tissue concentrations and in vitro production of P4. We inferred that increased P4 concentrations at the ovarian-uterine level in pLH-treated cows could have promoted embryo development and increased pregnancy per AI.

GnRH agonists: Updating fixed-time artificial insemination protocols in sows.

Protocols for fixed-time artificial insemination (FTAI) in swine reproduction can help increase genetic improvement and production efficiency. Different gonadotropin-releasing hormone (GnRH) agonists have been developed to gain better control of follicular development, timing, and ovulation quality; therefore, they have been extensively used in FTAI protocols. This literature review resumes the most important characteristics of the physiology of follicular development and ovulation in sows, followed by a discussion about the hormonal alternatives available to induce ovulation (human chorionic gonadotropin, hCG; porcine luteinizing hormone, LH and GnRH agonists). Also, ovulation induction failures with GnRH agonists are described. Finally, current FTAI protocols with GnRH agonists are resumed and discussed. FTAI with GnRH agonists has proven to be an efficient, successful reproductive protocol that can be implemented in pig farms due to better knowledge of an endocrine system that regulates follicular development and ovulation and increased availability of several GnRH agonists that allow more efficient reproductive swine programs.

Does a single fixed-time insemination in weaned sows affect gestation and lactation length, and piglet performance during lactation compared with multiple insemination protocols in a commercial production setting?

This study evaluated the effect of different single fixed-time artificial insemination (FTAI) protocols on gestation length, farrowing distribution (synchrony), and piglet performance. In Study 1, 866 sows were assigned to two groups: Multiple AI (n = 484) – multiple artificial insemination (AI) at 24 h intervals during estrus; and FTAI+MultAI (n = 382) – OvuGel® 96 h post-weaning and single FTAI 22–24 h later. In Study 2, FTAI protocols were retrospectively analyzed: pLH – sows received 2.5 mg (Exp.1, n = 184) or 5 mg (Exp.2, n = 362) of porcine luteinizing hormone (pLH) at estrus onset, and single FTAI 24 h later. The FTAI+MultAI resulted in shorter gestation length (P < 0.01) and greater synchrony of farrowing on days 114–115 of gestation (P < 0.01) than Multiple AI. Longer lactation length (21.2 vs. 20.9 d; P = 0.02) and greater piglet weaning weight (5378.9 ± 73.1 vs. 5153.2 ± 74.4 g; P = 0.03) for FTAI+MultAI compared to Multiple AI were observed. In Study 2, the gestation length based on the first AI record was shorter for FTAI and resulted in greater synchrony of farrowing on days 114–115 of gestation, compared with Multiple AI (P ≤ 0.05). Gestation length did not differ between groups (P > 0.05) when measured based on most probable AI responsible for fertilization (ultrasound evaluation). In conclusion, FTAI resulted in shorter gestation length and greater farrowing synchrony compared with Multiple AI. Longer lactation length and greater piglet weaning weight were observed for FTAI compared with multiple insemination protocols.

Successful Laparoscopic Oviductal Artificial Insemination in the Endangered Tsushima Leopard Cat (Prionailurus bengalensis euptilurus)

Simple SummaryThe Tsushima leopard cat is an endangered wild felid that lives solely on Tsushima Island, Nagasaki, Japan. Approximately, only 100 Tsushima leopard cats can be found in the wild, and there are concerns that the population will further reduce due to habitat degradation and traffic accidents. In 1994, Japan’s Ministry of the Environment (MOE) developed a conservation and breeding project for the Tsushima leopard cat. The MOE is working with the Japanese Association of Zoo and Aquariums for ex situ conservation of this species. However, considering genetic diversity, it is difficult to conduct captive breeding programs using only natural breeding; hence, assisted reproductive technologies are required. This study aimed to breed Tsushima leopard cats using artificial insemination (AI) by depositing sperms into the oviducts. Ovulation was artificially induced in two females, laparoscopically inseminated with fresh sperms into the oviducts. The pregnancies were monitored via fecal levels of progestogens and radiography. One female had spontaneous delivery of a female kitten 66 days post-AI. This is the first successful case of AI in a Tsushima leopard cat.The Tsushima leopard cat (Prionailurus bengalensis euptilurus) is an endangered wild felid that lives solely on Tsushima Island, Nagasaki, Japan. Japan’s Ministry of the Environment is working with the Japanese Association of Zoo and Aquariums for ex situ conservation of this species. However, considering genetic diversity, it is difficult to conduct captive breeding programs by natural breeding alone; hence, assisted reproductive technologies are required. This study aimed to breed Tsushima leopard cats, which otherwise cannot be paired, using laparoscopic oviductal artificial insemination (AI). Female Tsushima leopard cats (female 1, aged 7 years; female 2, aged 6 years) were treated with 200 IU equine chorionic gonadotropin, followed by administration of 1000 IU porcine luteinizing hormone (pLH) after 96 h to induce ovulation. Laparoscopic AI was performed 32 h post-pLH administration. Females 1 and 2 were inseminated in the oviduct with 2.4 × 10⁶ and 3.3 × 10⁶ motile spermatozoa, respectively, collected from two males. Pregnancy was confirmed by radiography 45 and 51 days post-AI in females 1 and 2, respectively; one fetus was found in female 2. Moreover, female 2 had spontaneous delivery of a female kitten 66 days post-AI. This is the first successful case of AI in a Tsushima leopard cat.

Benefits of synchronizing ovulation with porcine luteinizing hormone (pLH) in a fixed time insemination protocol in weaned multiparous sows

Materials and methods: Multiparous sows were randomly assigned to treatment at weaning. From weaning, twice-daily boar exposure facilitated estrus detection. Untreated control sows (n = 150; CONT) were artificially inseminated at least twice. Treated sows (n = 168; LUT) received 5 mg pLH intramuscularly concomitant with the first detection of standing heat. To coincide with the normal working day, sows in estrus in the morning were inseminated at 24 and 30 hours (am and pm), while sows in estrus in the afternoon were inseminated at 24 and 42 hours (pm and am), after pLH administration.

Human chorionic gonadotrophin does not induce ovulation in the tammar wallaby

We investigated whether human chorionic gonadotrophin (hCG), rather than porcine luteinising hormone (pLH), could induce ovulation in the tammar wallaby (Notamacropus eugenii) after priming with pregnant mare serum gonadotrophin (PMSG). pLH is effective, but is more expensive and supply is problematic. On Day 0 pouch young were removed (RPY) and 5mgkg−1 of bromocriptine mesylate was administered to all non-pregnant wallabies (n=12) to reactivate an oestrous cycle. On Day 20 RPY each female received 20IU PMSG i.m. and three days later (Day 23 RPY), the Single-Dose group (n=6) received 500IU of hCG suspended in oil at 0900 hours, and the Multi-Dose group (n=6) received 500IU of hCG at 0900, 1230 and 1630 hours. From Day 25 to Day 28 RPY copulatory plugs were observed in the Single-Dose (n=4 of 6; Day 25.3 to 28.3 RPY) and Multi-Dose (n=5 of 6; Day 25 to 28 RPY) groups. Autopsy on Day 31 RPY revealed that all females had reactivated, and despite all having large preovulatory follicles (4–6mm) in both ovaries, no ovulations or embryos were observed. We conclude that hCG is not an effective source of exogenous LH for the induction of ovulation in the tammar wallaby.

Comparison of Single, Fixed-Time Artificial Insemination in Gilts Using Two Different Protocols to Synchronize Ovulation.

Simple SummaryIt is necessary to have a consistent supply of service-ready gilts available to incorporate into each batch of breeding sows. Techniques to manipulate the timing of estrus and possibly the timing of ovulation in gilts are helpful in order to achieve this goal. This study investigated two different techniques for inducing ovulation to allow fixed-time artificial insemination (FTAI) protocols in gilts and compared results with gilts bred when observed in standing heat following cessation of daily altrenogest treatment. Pubertal gilts (n = 180) were assigned to one of three treatment groups. Group 1 gilts (LUT, n = 62) were induced to ovulate utilizing intramuscular injections of equine chorionic gonadotropin followed by porcine luteinizing hormone, and bred using a single FTAI. Group 2 gilts (TRI, n = 61) were induced to ovulate by intravaginal deposition of triptorelin acetate and were bred by a single FTAI. Group 3 gilts (CON, n = 57) were observed for estrus and bred twice (24 h apart) using artificial insemination (AI). LUT and TRI gilts completed farrowing in a smaller window of time compared to CON gilts; however, they also tended to have poorer reproductive performance. LUT and TRI piglets were 80 g and 64 g, respectively, heavier at weaning than CON piglets. Results indicate that FTAI might be useful as a means of minimizing the range in lactation length in a farrowing batch. However, modifications of the protocols may be required to ensure optimum farrowing rates and litter size.In order to efficiently have a consistent supply of service-ready gilts available to incorporate into each batch of breeding sows, it is necessary to manipulate the timing of estrus and possibly the timing of ovulation of gilts. Estrus can be synchronized by the withdrawal of altrenogest after at least 14 days of treatment. It is possible that protocols developed to induce ovulation, and therefore allow fixed-time artificial insemination (FTAI), can improve the predictability of gilt breeding. This study investigated the effect of two FTAI protocols in gilts on reproductive performance and timing of farrowing and piglet weaning weight compared to gilts bred based on signs of estrus after cessation of altrenogest. Puberty was induced in gilts, followed by treatment with altrenogest. Following altrenogest withdrawal, 180 gilts were assigned to one of three treatment groups. Group 1 gilts (LUT, n = 62) were treated with 600 IU equine chorionic gonadotropin 24 h after altrenogest withdrawal and 5 mg porcine luteinizing hormone (pLH) 80 h later, followed by a single FTAI 36 h after pLH. Group 2 gilts (TRI, n= 61) received 2 mL of a gonadotropin-releasing hormone agonist, triptorelin acetate, intravaginally 6 d after altrenogest withdrawal and were bred by a single FTAI 24 h later. Group 3 gilts (CON, n = 57) were observed for estrus and bred twice by AI, 24 h apart. LUT and TRI gilts farrowed closer together (2.4 ± 1.6 and 2.9 ± 1.2 d(days), respectively) compared to CON gilts (4.5 ± 3.3 d). Piglets in LUT were 80 g (p < 0.001) heavier and piglets in TRI were 64 g (p < 0.05) heavier at weaning than CON piglets, when controlling for birth weight. Results indicate that FTAI might be useful as a means of minimizing the time from the first to the last gilt farrowing in a breeding batch of gilts. However, modifications of the protocols may be required to ensure optimum farrowing rates and litter size.

Combining Fixed-Time Insemination and Improved Catheter Design in an Effort to Improve Swine Reproduction Efficiency

Simple SummaryConventional practice is to breed sows by artificial insemination (AI) at least twice using approximately three billion sperm per insemination. Sows may be bred only once using the technique of fixed-time artificial insemination (FTAI) if ovulation is predictable. This research explored the use of combining a single fixed-time artificial insemination (FTAI) and an alternative insemination catheter design that reportedly reduces semen backflow in order to reduce semen dosage and maintain reproduction efficiency. The FTAI technique used in this study involved two hormone treatments, 80 h apart, after weaning followed by a single insemination. The two catheters used in this study were a conventional foam-tipped insemination catheter and a Gedis catheter, which is designed to be completely inserted into the vagina of the sow. The semen is enclosed along the length of the catheter and held in place by a gel cap that melts when inserted into the cervix. Sows were assigned to the following treatments: Group 1 (n = 135), bred twice with a conventional catheter and a standard semen dose of approximately three billion sperm in 80 mL; Group 2 (n = 123), FTAI with conventional catheter and a standard semen dose; Group 3 (n = 127), FTAI with Gedis catheter and a standard semen dose; Group 4 (n = 126), FTAI with Gedis catheter and a reduced semen dose with approximately one billion sperm. The farrowing rates were 81.6%, 77.7%, 74.0%, and 62.7% for Groups 1 to 4, respectively. Litter sizes of Group 3 and Group 4 were smaller than Group 1. Overall, the combination of Gedis catheter and FTAI resulted in decreased reproductive performance that outweighed the value of using less semen.Conventional practice is to breed sows by artificial insemination (AI) at least twice using approximately three billion sperm per insemination upon estrus at standing heat. This research explored the use of combined technologies, including fixed-time insemination (FTAI) and an alternative catheter design that reportedly reduces semen backflow, in order to reduce the number of inseminations and the semen dosage and maintain reproductive efficiency. The FTAI technique used in this study was to inject I.M. 600 IU equine chorionic gonadotropin (eCG) at weaning and 5 mg porcine luteinizing hormone (pLH) to stimulate ovulation 80 h later, followed by a single insemination 36 h after the pLH injection. The two catheters used in this study were a conventional foam-tipped insemination catheter and a Gedis catheter. The Gedis catheter is designed to be completely inserted into the vagina. The semen is enclosed along the length of the rod and held in place by a gel cap that melts when inserted into the cervix. Sows were assigned to the following treatments: Group 1 (n = 135), bred twice with a conventional catheter and a standard semen dose of approximately three billion sperm in 80 mL; Group 2 (n = 123), FTAI with conventional catheter and a standard semen dose; Group 3 (n = 127), FTAI with Gedis catheter and a standard semen dose; Group 4 (n = 126), FTAI with Gedis catheter and a reduced semen dose with one billion sperm. The farrowing rates were 81.6%, 77.7%, 74.0%, and 62.7% for Groups 1 to 4, respectively. The likelihood of farrowing was lower for Group 3 and Group 4 compared to Group 1 (odds ratio (OR) = 0.57; p = 0.08 and OR = 0.35; p = 0.001, respectively). Likewise, litter size of Group 3 and Group 4 was smaller than Group 1 (p = 0.006 and p = 0.04, respectively). Overall, the combination of Gedis catheter and FTAI resulted in decreased reproductive performance that outweighed the value of using less semen.

Responsiveness of the cheetah (Acinonyx jubatus) ovary to exogenous gonadotropins after preemptive oral progestin treatment

Control of ovarian function in cheetahs is sub-optimal, which currently limits the integration of assisted reproductive techniques into the genetic management of that endangered species. The objective of this study was to determine the effect of preemptive progestin treatment on the quality of ovarian responses after exogenous gonadotropin stimulation in cheetahs. Adult females received either 1) 200 IU equine chorionic gonadotropin (eCG) followed with 3,000 IU porcine luteinizing hormone (pLH) (intramuscular route) (n = 5; control group) or 2) similar eCG/pLH administration preceded by a 7-day treatment with oral progestin (0.1 mg/kg altrenogest; ALT group; n = 7). At 42 h post-pLH administration, a series of metrics was assessed via laparoscopy (number of follicles ≥ 2 mm, number of corpora lutea, oviduct and uterine cornua diameter and overall vascularization). Concentrations of fecal estradiol, progesterone and glucocorticoid metabolites (FEM, FPM, and FGM, respectively) were measured by enzyme immunoassay for 3 wk before ALT treatment (Period 1), 7 d during ovarian suppression period (Period 2), throughout eCG/LH treatment and laparoscopy (Period 3), and 6 wk following laparoscopy (Period 4). Overall, nine out of 12 cheetahs (4/5 in control and 5/7 ALT group) had freshly-formed corpora lutea at the time of laparoscopy. Mean follicle and corpora lutea numbers in the control versus ALT group were not different (P > 0.05). Overall measurements and vascularization scores also did not differ (P > 0.05) among groups. FEM average concentrations increased (P ≤ 0.05) in response to eCG for the ALT-treated females between Periods 2 and 3 and were sustained during Period 4. However, FEM average concentrations did not vary (P > 0.05) for control females throughout Periods 1–4. Post-ovulatory FPM average concentrations (Period 4) did not differ (P > 0.05) between the ALT-treated females and controls. FPM average concentration from both groups increased in Period 4 compared to Periods 1–3 (P ≤ 0.05). Females receiving the ALT treatment also had lower (P ≤ 0.05) FGM metabolite average concentrations than control females during ovarian suppression (suggesting adrenal suppression). Collective results suggest that ovarian response to gonadotropin treatment in the cheetah was improved following oral progestin administration due to the normative increase in estradiol following stimulation for these females compared with control. This treatment should lead to more effective timed assisted reproduction procedures for this species.

Reproductive performance of fixed-time artificial insemination in swine and factors for the technology success

ABSTRACT: Fixed-time artificial insemination (FTAI) is a reproductive technology that aids in obtaining an appropriate time to perform single artificial insemination (AI), thus reducing the number of inseminations per sow bred. FTAI protocols can either be based on estrus detection or day of weaning, aiming to synchronize ovulation using ovulation inducers. The protocols involving estrus detection usually employ porcine luteinizing hormone (pLH) as an inducer and, in general, satisfactory reproductive performance is observed. For protocols based on weaning day, the main hormone used is analog of gonadotropin-releasing hormone such as triptorelin and buserelin. Regardless of the protocol, the number of piglets born is usually not affected by FTAI. However, a possible compromise in the farrowing rate should be considered. The FTAI in gilts requires progestogen treatment for estrus synchronization, increasing the labor requirement and cost of protocol. Some of the benefits of FTAI are a reduced number of semen doses required, advantage of planning the breeding time and; consequently, optimizing labor involved. However, the limitations include a slight reduction in the fertility index due to the compromised farrowing rate in some cases, costs incurred by following the protocol, and difficulty in measuring all the conceptual benefits under commercial conditions. The aim of this review is to approach the reproductive performance of the current protocols of FTAI, considering the benefits and limitations of this technology in swine production.

SUCCESSFUL LAPAROSCOPIC OVIDUCTAL ARTIFICIAL INSEMINATION IN THE CLOUDED LEOPARD (NEOFELIS NEBULOSA) IN THAILAND.

Captive breeding of clouded leopards (Neofelis nebulosa) is challenging because of mating incompatibility, high incidence of teratospermia in males, and inconsistent ovulation patterns in females. Assisted reproductive techniques, therefore, are necessary to overcome these issues and maintain the genetic diversity in the captive population. The objective was to use laparoscopic oviductal artificial insemination (AI) to breed genetically valuable females (n = 4; aged 4.5-5 yr) that were unsuccessfully paired. Fecal hormone metabolites (estrogen and progesterone) were extracted and measured by enzyme immunoassay for monitoring of ovarian activity 45 days before and 65 days after laparoscopic AI. For timed insemination, females were injected with 200 IU equine chorionic gonadotropin and 1,000 IU porcine luteinizing hormone (pLH) at the 82-hr interval. Ovarian assessment was performed by laparoscopy 44 hr after pLH administration. One nulliparous female out of four presented two ovulation sites on each ovary. The single female that had ovulated was inseminated with chilled semen collected from two males (8 × 106 and 2.7 × 106 motile spermatozoa, respectively, in each oviduct). A significant increase in fecal progesterone concentrations was observed after AI with a concentration peak (500 μg/g dry feces) detected on day 24 after pLH injection, which was then sustained for more than 45 days after the pLH injection. The delivery of two cubs occurred on day 92 after pLH. Microsatellite marker analysis determined that both cubs were sired by the same male. This is the first report of a successful oviductal AI in the clouded leopard.

Single fixed-time artificial insemination in gilts and weaned sows using pLH at estrus onset administered through vulvar submucosal route

This study evaluated the use of a single fixed-time artificial insemination (FTAI) in gilts and weaned sows using 2.5 mg of porcine luteinizing hormone (pLH) administered through vulvar submucosal route, at the onset of estrus. In experiment 1 (Exp.1), 318 pubertal gilts were assigned to two groups: control-G—no hormonal application and artificial inseminations (AIs) at 12, 36, and 60 hours after the onset of estrus if they were still in standing estrus; and FTAI-G—use of pLH at the onset of estrus and a single FTAI 12 hours later. In experiment 2 (Exp. 2), 309 weaned sows were assigned to three groups: Control-S—no hormone application and AIs at 0, 24, and 48 hours after the onset of estrus if they were still in standing estrus; FTAI-NH—no hormone application and a single FTAI at 24 hours after the onset of estrus, and FTAI-pLH—use of pLH at the onset of estrus and a single FTAI 24 hours later. Transabdominal real time B-mode ultrasonography was performed to determine whether the insemination had been performed within 24 hours before ovulation, considered as the optimal interval. In Exp. 1, ultrasound evaluation (12-hour intervals) was carried out to determine the interval between the onset of estrus and ovulation. In both experiments, 2 × 109 sperm cells in 80 mL were used to perform cervical and postcervical deposition of semen in gilts and sows, respectively. Compared with control-G, FTAI-G gilts had shorter (P < 0.05) duration of estrus (57.7 vs. 61.2 hours) and interval between the onset of estrus and ovulation (36.3 vs. 42.3 hours). The adjusted farrowing rate (AFR) was lower (P < 0.05) in FTAI-G (86.0%) compared with control-G (93.5%), but total piglets born (TPB) did not differ between these groups (12.3 vs. 12.5 piglets). Within the FTAI-G group, the AFR was lower (P < 0.05) in the presence (50.0%) than in the absence (94.9%) of semen backflow during AI. Also in the FTAI-G group, the insemination outside the optimal interval reduced (P < 0.05) the TPB (10.5 vs. 12.9 piglets) in comparison with gilts inseminated within the optimal interval. In Exp. 2, there were no differences in the AFR (Control-S: 94.1%; FTAI-NH: 86.1%; FTAI-pLH: 88.0%) and TPB (Control-S: 12.8; FTAI-NH: 12.7, and FTAI-pLH: 12.0 piglets) among treatments. The presence of semen backflow reduced (P < 0.05) the TPB in FTAI-pLH and FTAI-NH sows. In the FTAI-pLH, a single insemination performed too late relative to ovulation reduced the AFR (P < 0.05) compared with sows inseminated within the optimal interval. In conclusion, 2.5-mg pLH applied at the vulvar submucosa at the onset of estrus advances the ovulation in gilts, but a single FTAI performed 12 hours later reduces the farrowing rate. A single FTAI performed at 24 hours after the onset of estrus in weaned sows does not affect their reproductive performance, regardless of pLH application.

Recent advancements in the hormonal stimulation of ovulation in swine.

Induction of ovulation for controlled breeding is available for use around the world, and conditions for practical application appear promising. Many of the hormones available, such as human chorionic gonadotropin (hCG), gonadotropin-releasing hormone (GnRH) and its analogs, as well as porcine luteinizing hormone (pLH), have been shown to be effective for advancing or synchronizing ovulation in gilts and weaned sows. Each of the hormones has unique attributes with respect to the physiology of its actions, how it is administered, its efficacy, and approval for use. The timing for induction of ovulation during the follicle phase is critical as follicle maturity changes over time, and the success of the response is determined by the stage of follicle development. Female fertility is also a primary factor affecting the success of ovulation induction and fixed time insemination protocols. Approximately 80%–90% of female pigs will develop mature follicles following weaning in sows and synchronization of estrus in gilts. However, those gilts and sows with follicles that are less developed and mature, or those that develop with abnormalities, will not respond to an ovulatory surge of LH. To address this problem, some protocols induce follicle development in all females, which can improve the overall reliability of the ovulation response. Control of ovulation is practical for use with fixed time artificial insemination and should prove highly advantageous for low-dose and single-service artificial insemination and for use with frozen-thawed and sex-sorted sperm.

Porcine luteinising hormone given at oestrus onset by intramuscular route does not advance ovulation in gilts

This study aimed to evaluate the use of porcine luteinising hormone (pLH) given at oestrus onset in gilts to synchronise ovulation. A total of 120 gilts (40/treatment) were assigned in three treatments: control - application of placebo by intramuscular (i.m.) route at oestrus onset; pLH2.5 - application of 2.5mg of pLH by i.m. route at oestrus onset; pLH5 - application of 5mg of pLH by i.m. route at oestrus onset. On average, the interval onset of oestrus to ovulation did not differ (P&gt;0.05) among treatments (control - 28.7±1.6h; pLH2.5 - 28.2±1.6h; pLH5 - 27.5±1.6h). The frequency distribution of gilts ovulated in different moments after oestrus detection was not affected (P&gt;0.05) by the treatment. In conclusion, the use of 2.5mg or 5mg of pLH given at oestrus onset in gilts by i.m. route does not advance and synchronises the interval onset of oestrus to ovulation.

163 RE-ESTABLISHING REPRODUCTIVE CYCLICITY IN A FEMALE ASIAN SMALL-CLAWED OTTER FOLLOWING SUPRELORIN-INDUCED CONTRACEPTION: A CASE STUDY

Asian small-clawed otters (ASCO) are a popular species to exhibit in zoological institutions globally, and are found in managed populations in the United States, Europe, Asia, and Australia. Captive breeding of these otters is integral to population sustainability, with management programs using pedigree analyses to make specific breeding recommendations to ensure long-term genetic viability. Because of the familial social structure of ASCO and limited space within zoos, physical separation of animals is not always possible for temporary breeding prevention, and short-term contraception may be preferred. In US zoos, Suprelorin (deslorelin; Virbac Australia, Milperra, Australia), a GnRH agonist, has been recommended for contraception of female carnivores, due to its small implant size and lack of side effects often associated with hormone-based contraceptives. However, the duration of reproductive suppression with Suprelorin may be excessive and reversibility (i.e. resumption of cyclicity, ovulation, and pregnancies) in implanted females across a range of species has been variable, unpredictable, and prolonged. Since 2006, 49 female ASCO have been implanted with Suprelorin at least once, and, of these, only two females have shown confirmed reversibility with pregnancies. No ASCO females implanted more than once have thus far exhibited reversibility (personal communication, AZA Wildlife Contraceptive Center, St. Louis, MO, USA). In this case study, fecal hormone monitoring of one female ASCO implanted with Suprelorin three times (Dec 2007, Jan 2009, Mar 2010), showed a lack of ovarian cyclicity and ovulation during the three years since her last implant. In an attempt to induce ovarian follicular growth and ovulation, this female was injected (IM) with exogenous gonadotropins (100 IU of equine chorionic gonadotropin followed 80 h later with 3000 IU of porcine luteinizing hormone). Fecal progesterone monitoring confirmed ovulation followed by a 76 day pseudopregnancy, with temporal characteristics similar to those previously reported in naturally cycling ASCO (Bateman et al. 2009 Zoo Biol. 28, 107–126). Following the induced pseudopregnancy and ~55 days of basal progestin levels, this female was observed breeding with a cohabitating ASCO male. Fecal hormone monitoring revealed subsequent ovulation and the occurrence of another pseudopregnancy of normal duration. These preliminary findings suggest that exogenous gonadotropin treatment may be useful for promoting resumption of normal ovarian cyclicity and ovulatory responses in ASCO following prolonged reproductive suppression with Suprelorin.

Gonadotropin-induced puberty does not impair reproductive performance of gilts over three parities.

The aim of this study was to evaluate the reproductive performance of three parities of gilts treated or not treated with gonadotropin to induce puberty. Sixty gilts received 600 IU of equine chorionic gonadotropin (eCG) followed by 2.5 mg of porcine luteinizing hormone (LH) 72 h later. Fifty-nine other gilts were exposed only to a mature boar for 15 min twice daily. Artificial insemination (AI) was performed at 0, 12 and 24 h after the detection of oestrus, and gestation was confirmed by ultrasound after 35 days. Sows were inseminated at the first post-weaning oestrus. The total numbers of piglets born, piglets born alive, stillborn, mummified foetuses, as well as pregnancy and farrowing rates were evaluated for each of the three parities. Culling rates, farrowing intervals and weaning-to-oestrous intervals (WEI) were also analysed. Mean age at puberty and oestrous manifestation were not significantly different between treatments (p = 0.0639; 179.20 ± 17.52 compared with 173.96 ± 16.94, 91.66% compared with 94.92%) across the experimental period. However, females that underwent puberty induction showed modest increases both in the number of total pigs born and in the number of piglets born alive. In conclusion, puberty induction through exogenous gonadotropin administration in field conditions did not induce a more concentrated first oestrous manifestation, but trended to a modest increase in the number of pigs born alive in the first parity and a reduced culling rate during the first gestation.

Ovulation induction and artificial insemination of a captive polar bear (Ursus maritimus) using fresh semen.

In 2008, polar bears were listed as a species threatened with extinction by the U.S. Endangered Species Act. Unfortunately, reproductive success has been poor despite breeding recommendations for almost every reproductively viable bear by the Species Survival Plan. Assisted reproductive technologies could complement breeding efforts by overcoming the challenges of behavioral incompatibilities and deficiencies, facilitating genetic management and increasing cub production. The goal of this study was to artificially inseminate a female polar bear after inducing ovarian activity and ovulation with exogenous hormones (equine chorionic gonadotropin and porcine luteinizing hormone). Fresh semen collected from an adult male via electroejaculation/urethral catheterization was used for the insemination. Fecal steroid monitoring indicated that the female ovulated following the exogenous hormone treatment. Progestin concentrations increased in late summer, at the time implantation was expected to occur; however, no cubs were produced. To the authors' knowledge, this is the first report of ovulation induction and artificial insemination in a polar bear.

Precision-cut luteal slices: A promising approach for studying luteal function in pigs

The study was aimed to validate the precision-cut luteal slices to investigate porcine luteal function. Corpora lutea (CLs) were cut into 180-μm thick slices using Krumdick Tissue Slicer. The viability, tissue structure and steroidogenic acute regulatory protein (STAR) expression in the luteal slices did not differ between the beginning and the end of the 24-h incubation period. The luteal progesterone secretion showed a time- and dose-dependent response to porcine luteinizing hormone. The effects of prostaglandin F2α and 17β-estradiol on progesterone secretion by porcine luteal slices were comparable to the previously reported in vivo results of the CL microdialysis system in the pig.

Fixed-time post-cervical artificial insemination in sows receiving porcine luteinising hormone at oestrus onset

Fixed-time post-cervical artificial insemination (FTPCAI) allows a wider use of high indexing boars and a considerable reduction in labour requirements in swine production. The aim of this study was to evaluate fixed-time artificial insemination (FTAI) efficiency using different artificial insemination protocols and porcine luteinising hormone (pLH) to induce ovulation. A total of 597 weaned sows in which oestrus detection was performed once daily (08:00 am) was allocated to three groups: FTPCAI1 (n=199) – sows received a 5-mg (4ml) intramuscular injection of pLH at oestrus onset, and were inseminated 24h later; FTPCAI2 (n=199) – sows received 5mg of pLH and were inseminated at oestrus onset (0h) and 24h after; and MultPCAI (n=199) – sows did not receive pLH, and the first AI was performed at oestrus onset (0h) and repeated every 24h during oestrus. Homospermic doses (1.5×109 total sperm cells/50ml) were used in post-cervical artificial insemination (PCAI) in all the treatments. Hormonal treatment did not affect (P>0.05) the interval between oestrus onset and ovulation (overall 32.4h) and there were no differences (P>0.05) in farrowing rate (overall 91.6%) or litter size (overall 12.6 piglets born) among treatments. In sows treated with pLH at oestrus onset, a single PCAI with 1.5 billion sperm cells did not compromise reproductive performance compared with a double PCAI at 24h intervals.

Laparoscopic Oviductal Artificial Insemination Improves Pregnancy Success in Exogenous Gonadotropin-Treated Domestic Cats as a Model for Endangered Felids1

Artificial insemination (AI) in cats traditionally uses equine chorionic gonadotropin (eCG) and human chorionic gonadotropin (hCG) to induce follicular development and ovulation, with subsequent bilateral laparoscopic intrauterine insemination. However, long-acting hCG generates undesirable secondary ovulations in cats. Uterine AI also requires relatively high numbers of spermatozoa for fertilization (~8 × 10(6) sperm), and unfortunately, sperm recovery from felids is frequently poor. Using short-acting porcine luteinizing hormone (pLH) instead of hCG, and using the oviduct as the site of sperm deposition, could improve fertilization success while requiring fewer spermatozoa. Our objectives were to compare pregnancy and fertilization success between 1) uterine and oviductal inseminations and 2) eCG/hCG and eCG/pLH regimens in domestic cats. Sixteen females received either eCG (100 IU)/hCG (75 IU) or eCG (100 IU)/pLH (1000 IU). All females ovulated and were inseminated in one uterine horn and the contralateral oviduct using fresh semen (1 × 10(6) motile sperm/site) from a different male for each site. Pregnant females (11/16; 69%) were spayed approximately 20 days post-AI, and fetal paternity was genetically determined. The number of corpora lutea (CL) at AI was similar between hormone regimens, but hCG increased the number of CL at 20 days post-AI. Numbers of pregnancies and normal fetuses were similar between regimens. Implantation abnormalities were observed in the hCG group only. Finally, oviductal AI produced more fetuses than uterine AI. In summary, laparoscopic oviductal AI with low sperm numbers in eCG/hCG- or eCG/pLH-treated females resulted in high pregnancy and fertilization percentages in domestic cats. Our subsequent successes with oviductal AI in eCG/pLH-treated nondomestic felids to produce healthy offspring supports cross-species applicability.