Unfertilized Ova Research Articles

Parthenogenesis, identity, and value.

Parthenogenesis is a form of asexual reproduction in which a gamete (ovum or sperm) develops without being fertilized. Tomer Jordi Chaffer uses parthenogenesis to challenge Don Marquis'future-like-ours (FLO) argument against abortion. According to Marquis, (1) what makes it morally wrong to kill us is that it would deprive us of a possible future that we might come to value-a future "like ours" (FLO) and (2) human fetuses are numerically identical to any adult human organism they may develop into, and thus have a FLO. Chaffer contends that if human ova are capable of parthenogenesis, then they would have a FLO, which contraception may deprive them of, but contends this is absurd. Bruce P. Blackshaw challenges Chaffer, contending sexually fertilized embryos are not identical to unfertilized ovum, but this would yield a more absurd implication, that fertilization deprives an ovum of a FLO! Here I show Marquis'account of identity rules out both Chaffer's and Blackshaw's accounts.

Evaluasi Pengaruh Level Kandungan Protein dalam Pakan terhadap Respons Super Ovulasi: Kajian Meta-Analisis

The purpose of this study was to evaluate and analyze the effect of the level of protein content in feed on the superovulation response in cattle through a meta-analysis study. The meta-analysis study was performed with OpenMEE application and used scientific articles from international journals according to the topic of the study. The superovulation parameters used in this study were the number of corpus luteum, the number of collected embryos, the number of viable embryos, degenerated embryos, unfertilized embryos, and the proportion of viable embryos. The results of a meta-analysis of 37 research results in 20 scientific articles showed feed protein level had a significant effect on increasing the number of corpus luteum, collected embryos, viable embryos, and the proportion of viable embryos. Meanwhile, the feed protein level had no significant effect on degenerated embryos and unfertilized ovum. It could be concluded that an increase in feed protein content had a positive effect on several reproductive quality parameters.
 Key words: cow, embryo, feed, protein, reproduction, superovulation

Thyroid hormone triiodothyronine does not protect ovarian reserve from DNA damage induced by X-ray and cisplatin.

Cancer therapy can induce premature ovarian insufficiency, necessitating methods for preserving fertility in female cancer patients. However, the only accepted clinical practice for doing so is cryopreservation of embryos, unfertilized ova, and ovarian tissue, despite potential options such as in vitro maturation of follicles. Therefore, considerable interest has arisen in fertoprotective agents, with research on rat ovarian granulosa cells suggesting that triiodothyronine (T3) regulates an anti-apoptosis mechanism that protects the ovarian reserve from paclitaxel-induced DNA damage. In this study, we used postnatal day 5 mouse ovary to confirm the existence of T3 thyroid hormone receptor (THR), as well as to investigate the potential protective effects of T3 against cisplatin- and X-ray-induced apoptosis. We also tested the potential anti-apoptotic effect of T3 in the breast cancer cell line MDA-MB-231. We treated cultured mouse ovaries with varying concentration of T3 and 4μM cisplatin and 0.2Gy X-ray. Real-time PCR, histological analysis, immunoblot analysis, and immunofluorescence were performed to assess the potential anti-apoptotic effects of T3. We confirmed that THR alpha and beta are expressed in the mouse ovary. T3 (0.1, 1, 10, 100nM, and 1µM) does not protect ovarian reserve from cisplatin- or X-ray-induced apoptosis or DNA damage. Similarly, it does not protect mouse granulosa cells and MDA-MB-231 cells from cisplatin- or X-ray-induced apoptosis. Our findings suggest that T3 is ineffective as a fertoprotective agent, and its candidacy as a potential agent to preserve fertility should be reconsidered.

Influence of breed on the quality of in vivo produced embryos from Boran and Holstein Friesian cross dairy breed in Ethiopia

The variation of the dairy breed can determine the success of bovine embryo transfer by influencing the quantity and quality of in vivo embryo production. In this experiment, output and quality of in vivo produced embryos using semen of progeny tested Holstein Friesian (HF) sire in Boran and HF*Boran F1 cross cows, and semen from purebred Boran sire in HF*Boran F1 cross and Boran cows were evaluated. Boran (n=18) and HF*Boran cross (n=18) breed donor dams were superovulated using a previously optimized follicular stimulating hormone (FSH) (Pluset®) dose regimen: 650 IU for HF*Boran cross and 250 IU for Boran breeds. Each cow was flushed on Day-7 post insemination and embryos were evaluated for their developmental stages and quality. Superovulatory response rates were 88.9% and 83.3%, respectively, for Boran and HF*Boran with no significant (P>0.05) breed differences. Total recovery rates were relatively lower (56.5%) in Boran compared to in HF*Boran (67.4%). The mean (±SE) embryo flush outputs were 6.5±0.8 for Boran and 6.9±0.7 forHF*Boran with no significant breed difference. Recovery of a transferrable embryo was significantly higher (68.0%; P<0.05) in HF*Boran dam inseminated with HF sire semen. Boran cows yielded a significantly higher (P<0.05) proportion of unfertilized ovum (57.6 %) irrespective of the sire breeds. Comparatively, a higher number of degenerated embryos were produced by HF*Boran cows. This study demonstrated that the presence of breed-related differences in both the quality and quantity of in vivo produced Bovine embryos.

Süperovulasyon Öncesi GnRH ve Kısa Süreli Progesteron Uygulamalarının Sığır Embriyolarının Sayısı ve Kalitesi Üzerine Etkisi

Bu çalışmada, insan koryonik gonadotropini (hCG) ve prostaglandin F2-alfa (PGF2α) ile östruslarının indüklenmesinden sonra gonadotropin salgılatıcı hormon (GnRH) ve kısa süreli progesteron uygulamasını takiben süperovulasyon uygulanan Holştayn donörler (modifiye protokol) ile klasik süperovulasyon uygulanan Holştayn donörlerden elde edilen embriyoların kalitesi ve sayısı karşılaştırıldı. Araştırmada, üç yaşlı 20 adet Holştayn ırkı inek iki eşit gruba ayrıldı. Grup I’ deki hayvanlara 11 gün ara ile iki defa kloprostenol enjeksiyonu yapıldı ve östrus sonrası takip eden siklusun 9. gününden itibaren, günde iki kez dört gün süresince azalan dozlarda FSH enjeksiyonları uygulandı. Beşinci ve 6. FSH enjeksiyonları ile birlikte kloprostenol uygulamaları yapıldı.Grup II’ deki hayvanlara ise siklusun herhangi bir döneminde hCG uygulaması yapıldı. Bundan 7 gün sonra kloprostenol enjeksiyonu yapıldı. Östrus sonrası devam eden siklusun 7. günü buserelin enjeksiyonu yapıldı. PRID siklusun 9. günü yerleştirildi ve siklusun 14. günü çıkarıldı (6. FSH enjeksiyonu sırasında). Grup II’de FSH ve kloprostenol uygulamaları siklusun 12-15. günlerinde Grup I’ deki gibi yapıldı. Tüm inekler son FSH enjeksiyonundan sonra 12 saat ara ile üç kez tohumlandı. İkinci suni tohumlama ile eş zamanlı olarak buserelin enjeksiyonu yapıldı. İkinci tohumlama sonrası 7. günde embriyolar toplandı, morfolojilerine bağlı kalite ve gelişim evreleri değerlendirildi. Bu çalışmada donör başına sırasıyla Grup I ve Grup II’ de toplam CL (8.50±1.90 ve 9.40±1.52), toplam oosit ve embriyo (7.5±2.03 ve 7.0±1.61), toplam embriyo (6.00±2.00 ve 6.10±1.47), transfer edilebilir embriyo ı (5.50±1.84 ve 5.50±1.59), dejenere embriyo (0.50±0.22 ve 0.60±0.40) ve unfertilize oosit sayıları (1.60±0.97 ve 0.90±0.31) tespit edildi. Sonuç olarak, iki protokol arasında embriyo sayısı ve kalitesi açısından istatistiksel olarak fark bulunamadı (P >0,05). Böylece çok sayıda donöre östrus takibine gerek duyulmadan sabit zamanlı süperovulasyona olanak sağladığından dolayı klasik süperovulasyon uygulamalarına alternatif bir yöntem olabileceği sonucuna varılmıştır.

12 Artificial insemination and embryo transfer results in ewes during a long daylength period

Sheep are polyestrous, short-day breeders (∼11h of daylength), and exhibit oestrus approximately every 16–17 days during the breeding season, usually in late September to late December in the Northern Hemisphere. Progressive sheep producers often use assisted reproductive techniques such as laparoscopic AI and ovarian hyper-stimulation, embryo collection, and embryo transfer (ET) to increase genetic gain, and strive to have early December lambs to target specific show markets, which dictates AI or ET during the late summer. This field trial compared pregnancy rates following AI or ET in July and August (∼14h of daylength) in southwest Virginia (36–38′12″ N). Ewes (AI, n=83; ET recipients, n=33) were synchronized using a modified Ovsynch protocol involving intravaginal progesterone implants for 14 days, prostaglandin F2α (intramuscular) 48h before expected oestrus, and PG600 (IM) and gonadotrophin-releasing hormone (intramuscular) 52 to 54h and 16h before AI, respectively. Ewes were subjected to AI (frozen/thawed semen) regardless of whether they displayed signs of oestrus, and ewes selected as embryo recipients were subjected to a similar protocol but instead received 1 or 2 embryos (based upon the number of viable embryos produced per embryo donor) 6 days following the AI of the embryo donors. Ovarian hyper-stimulation of the embryo donors (n=13) was enabled by twice-daily FSH injections [totalling 350–455IU of Folltropin V (10–13mL)] for the 4 days before AI. Six days following AI, embryos were recovered surgically from the embryo donors (n=13) and yielded an average (±s.e.m.) of 6.6±1.2 total ova, 4.7±1.1 transferable quality embryos, and 1.9±0.8 unfertilized ova per collection. Pregnancy was detected using transrectal ultrasonography at ∼30 days of gestation and the pregnancy rates were analysed using Chi-squared. There was a tendency (P=0.092) for more pregnancies to be established following ET (22/33; 66%) compared with AI (41/83; 49%). There was no statistical relationship between AI ewes or ET recipient ewes that became pregnant relative to whether they displayed signs of oestrus or not. Embryo transfer was a more successful approach to produce pregnancies in ewes compared with AI during long daylength periods in this field trial. Further, ova from hyper-stimulated embryo donor ewes experienced a very high fertilization rate. Future studies will evaluate the ova capability directly via laparoscopic aspiration of ovarian follicles and IVF and evaluation of hyper- and non-hyper-stimulated follicular waves (using transrectal ultrasonography) and endocrine dynamics during different long and short daylength periods. Extending the opportunity to generate embryos and offspring regardless of daylength will assist aggressive sheep producers in reaching their reproductive, financial, and genetic goals.

Facultative Parthenogenesis in a Diamondback Water Snake (<i>Nerodia rhombifer</i>)

Here we describe a case of facultative parthenogenesis in a diamondback water snake (Nerodia rhombifer). An adult female N. rhombifer kept in captivity produced four unfertilized ova, one stillborn and one live neonate in July 2016. The neonates had characteristic abnormalities in morphology and were determined as males, suggestive of a parthenogenetic event. Stillborn and live neonates along with the mother and four unrelated N. rhombifer were genetically screened at twelve microsatellite loci. Results confirmed that the reproductive event represented a case of facultative parthenogenesis. In light of previous reports of facultative parthenogenesis in the genera Nerodia and Thamnophis, these results suggest that this reproductive mode may be a widespread phenomenon in the Natricinae as a whole.

Parthenogenesis and developmental constraints.

The absence of a paternal contribution in an unfertilized ovum presents two developmental constraints against the evolution of parthenogenesis. We discuss the constraint caused by the absence of a centrosome and the one caused by the missing set of chromosomes and how they have been broken in specific taxa. They are examples of only a few well-underpinned examples of developmental constraints acting at macro-evolutionary scales in animals. Breaking of the constraint of the missing chromosomes is the best understood and generally involves rare occasions of drastic changes of meiosis. These drastic changes can be best explained by having been induced, or at least facilitated, by sudden cytological events (e.g., repeated rounds of hybridization, endosymbiont infections, and contagious infections). Once the genetic and developmental machinery is in place for regular or obligate parthenogenesis, shifts to other types of parthenogenesis can apparently rather easily evolve, for example, from facultative to obligate parthenogenesis, or from pseudoarrhenotoky to haplodiploidy. We argue that the combination of the two developmental constraints forms a near-absolute barrier against the gradual evolution from sporadic to obligate or regular facultative parthenogenesis, which can probably explain why the occurrence of the highly advantageous mode of regular facultative parthenogenesis is so rare and entirely absent in vertebrates.

Fertility Preservation Using GnRH Agonists: Rationale, Possible Mechanisms, and Explanation of Controversy.

The only clinically accepted method of fertility preservation in young women facing gonadotoxic chemo- and/or radiotherapy for malignant or autoimmune diseases is cryopreservation of embryos or unfertilized ova, whereas cryopreservation of ovarian tissue for future reimplantation, or in vitro maturation of follicles, and the use of gonadotropin-releasing hormone agonists (GnRHa) are still considered investigational, by several authorities. Whereas previous publications have raised the fear of GnRHa’s possible detrimental effects in patients with hormone receptor-positive breast cancers, recent randomized controlled trials (RCTs) have shown that it either improves or does not affect disease-free survival (DFS) in such patients. This review summarizes the pros and cons of GnRHa co-treatment for fertility preservation, suggesting 5 theoretical mechanisms for GnRHa action: (1) simulating the prepubertal hypogonadotropic milieu, (2) direct effect on GnRH receptors, (3) decreased ovarian perfusion, (4) upregulation of an ovarian-protecting molecule such as sphingosine-1-phosphate, and (5) protecting a possible germinative stem cell. We try to explain the reasons for the discrepancy between most publications that support the use of GnRHa for fertility preservation and the minority of publications that did not support its efficiency.

Simplified superovulation protocols in dromedary camels (Camelus dromedarius)

This study was conducted to develop simple superovulation protocols in dromedary camels. Using two commercial FSH products, a series of experiments was conducted to evaluate the effect of different superovulation protocols on ovarian response and embryo production. In experiment 1, camels in control group (n = 15) received 400 mg Folltropin-V in a traditional protocol (FSH diluted in saline and given twice daily in decreasing doses over 5 days) and camels in split-injection (2 doses 48 h apart) groups received either 400 (n = 16, first dose: 320 mg, second dose: 80 mg) or 200 mg (n = 16, first dose: 120 mg, second dose: 80 mg) of slow-release (SR) preparation of Folltropin-V [Folltropin-V diluted in hyaluronan (5 mg/mL) solution]. In experiment 2, camels in control group (n = 13) received 2000 IU Pluset in a traditional protocol and camels in split-injection groups received either 2000 (n = 14, first dose: 1600 IU, second dose: 400 IU) or 1000 IU (n = 16, first dose: 600 IU, second dose: 400 IU) of SR preparation of Pluset (Pluset diluted in hyaluronan solution). In experiment 3, camels received SR preparation of 200 mg Folltropin-V (n = 45, first dose: 120 mg, second dose: 80 mg) or 1000 IU Pluset (n = 42, first dose: 600 IU, second dose: 400 IU) in a split-injection protocol. In experiments 1 and 2, the mean number of ovulations, corpora lutea, transferable embryos and unfertilized ova were similar (P > 0.05) between groups. In experiment 3, there was no difference (P > 0.05) between SR preparations of Folltropin and Pluset on the number of ovulatory sized (≥9 mm) follicles (15.8 ± 0.9 vs 16.7 ± 0.9) and transferable embryos (5.0 ± 0.4 vs 5.2 ± 0.5). In conclusion, split-injection of SR preparation of FSH resulted in a similar superovulatory response compared to a traditional protocol and a similar superovulatory response can be achieved with SR preparations of Folltropin-V and Pluset in a split-injection protocol.

Alternative male morphs solve sperm performance/longevity trade-off in opposite directions.

Males pursuing alternative reproductive tactics have been predicted to face a trade-off between maximizing either swimming performance or endurance of their sperm. However, empirical evidence for this trade-off is equivocal, which may be due to simplistic assumptions. In the shell-brooding cichlid fish Lamprologus callipterus, two Mendelian male morphs compete for fertilization by divergent means: Bourgeois nest males ejaculate sperm, on average, about six times farther from the unfertilized ova than do parasitic dwarf males. This asymmetry is opposite to the usual situation, in which bourgeois males typically benefit from superior fertilization opportunities, suggesting that nest males' sperm should persist longer than dwarf male sperm. The assumed trade-off between sperm swimming performance and longevity predicts that, in turn, sperm of dwarf males should outperform that of nest males in swimming efficiency. Measurement of sperm performance and endurance reveals that dwarf male spermatozoa swim straighter initially than those of nest males, but their motility declines earlier and their velocity slows down more abruptly. Nest male sperm survives longer, which relates to a larger sperm head plus midpiece, implying more mitochondria. Thus, the trade-off between sperm performance and endurance is optimized in opposite directions by alternative male morphs. We argue that the relative success of alternative sperm performance strategies can be influenced strongly by environmental factors such as the time window between gamete release and fertilization, and the position of gamete release. This is an important yet little understood aspect of gametic adaptations to sperm competition.

Climatic factors affecting quantity and quality grade of in vivo derived embryos of cattle

The present study investigated the effects of climatic variables on the quality grade and quantity of in vivo derived cattle embryos in the Midwestern United States. Climatic information included greatest and least daily temperature, average daily wind speed and average temperature-humidity index for each of the 765 records. The response variables included the number of ovarian structures, viable embryos, quality grade 1 embryos, quality grade 2 embryos, quality grade 3 embryos, freezable embryos (sum of quality grade 1 and quality grade 2 embryos), transferable embryos (sum of quality grade 1–3 embryos), degenerate embryos and unfertilized ova. Measures for variables among the breeds of donors and sires grouped by geographical origin were compared. A negative effect of greater temperatures during the early embryonic development stage tended (P < 0.10) to be associated with a decrease in the quality of embryos recovered. Interestingly, the greater the Temperature-Humidity Index (THI) during the early ovarian antral follicular development stage 40–45 days prior to ovulation was associated with a tendency for greater numbers of total number of freezable and transferable embryos recovered per uterine flushing (P < 0.10). Increased wind speed at the early antral follicular phase 40–45 days prior to ovulation was associated with an increase in the percentage of quality grade 1 embryos recovered (P < 0.05). Wind speed during the estrous synchronization period was also associated with a lesser number of embryos recovered (P < 0.05). This retrospective study confirms that climatic variables have significant effects on the in vivo production of cattle embryos and that wind speed should be considered in future analyses of factors affecting embryo quality.

Effect of breed and follicular status on response to superovulation in South African goats

ABSTRACTThis study evaluated the effect of breed and follicular status of Boer and indigenous goats on response to superovulation and embryo yield. The oestrous cycles were synchronized with progestagen for nine days and superovulated with porcine follicle-stimulating hormone. Does were cervically inseminated and embryos were surgically flushed on day six following artificial insemination. The oestrous activity, ovarian response, embryo yield and quality, did not differ significantly between breeds. The number and size of follicles at the onset of superovulation treatment and during oestrus did not differ significantly between breeds. The follicles 2–3 mm, 4–5 mm and total number of follicles at the onset of superovulation treatment were positively correlated with the number of structures, embryos and transferrable embryos recovered. The number of medium follicles (4−5 mm) at the beginning of superovulation treatment increased the number of transferable embryos. The total number of follicles >6 mm at the onset of superovulation was positively correlated to the number of unfertilized ova. Although limited number of animals was used, the results suggest that breed has limited effect on superovulation response. Instead, the quality and yield of embryos are dependent on number and size of follicles present at the beginning of a superovulation treatment.

59 EFFECT OF PELLET VOLUME AND THAWING TEMPERATURE ON VITRIFICATION EFFICACY WITH DOMESTIC CAT SEMEN COLLECTED VIA URETHRAL CATHETERIZATION

Historically, semen banking in felids has required sample collection via electroejaculation followed by sperm freezing in straws over LN2 vapor. Recent modifications include urethral catheterization of males treated with α-2 agonists for semen recovery and vitrification of cat sperm by suspension in a sucrose-based cryomedium and direct pelleting into LN2. In combination, these latter methods greatly simplify semen cryopreservation in cats but protocols need to be optimized for applied usage. In the present study, our goal was to assess the effect of 2 variables—pellet volume and thawing temperature—on post-thaw sperm motility, acrosome status, and in vitro fertility. Semen was collected from 3 males (3 ejaculates/male) via urethral catheterization under dexmedetomidine-ketamine anaesthesia. Sperm were diluted in Feline Optimized Culture Medium (FOCM), centrifuged (8 min; 300 × g), and resuspended in a soy-lecithin-based vitrification medium (with 0.2 M sucrose). After a 5-min equilibration, sperm was vitrified in 2 volumes (20 or 30 µL) by direct pipetting into LN2. Sperm pellets were thawed in FOCM at 1 of 2 temperatures (37 or 55°C) and the 4 treatment groups (20 µL-37°C, 20–55, 30–37, 30–55) assessed for percentage of progressively motile and acrosome intact sperm. To assess sperm function, additional 30-µL pellets were thawed at 37 or 55°C, and recovered sperm were used to inseminate in vitro-matured domestic cat oocytes (n = 10–25/ejaculate). At 48 h post-insemination, oocytes and embryos were fixed (1% NBF). Hoechst fluorescent stain (#33342) was used to evaluate embryo cleavage and maturation status of unfertilized ova. Sperm motility and acrosomal integrity percentages were analysed by ANOVA, and oocyte cleavage proportions were analysed by chi-squared. Mean (± SEM) progressive sperm motility post-thaw did not differ (P &gt; 0.05) among treatments (38 ± 8, 34 ± 7, 41 ± 7, 32 ± 7% for 20 µL-37°C; 20–55, 30–37, and 30–55, respectively). Similarly, acrosomal integrity did not differ (P &gt; 0.05) among treatments (26 ± 4, 25 ± 4, 17 ± 3, 17 ± 2% for 20 µL-37°C, 20–55, 30–37, and 30–55, respectively). Oocyte cleavage proportions did not differ (P &gt; 0.05) between thawing temperatures for total inseminated oocytes but, after correcting for oocyte maturation status, was higher (P &lt; 0.01) for samples thawed at 55°C (60%, 67/112) compared with 37°C (39%, 52/133). In summary, although variations in pellet volume and thawing temperature had minimal effect on sperm motility or acrosome status immediately post-thaw, sperm function appeared to be enhanced when vitrified pellets were thawed at a higher temperature. In vitro fertility success (~60% embryo cleavage) is comparable to values reported by our laboratory with conventionally collected and frozen cat semen, suggesting these newer methods may be suitable for applied usage in felids. This study was funded by the Institute of Museums and Library Services.

Embryo yield in dairy cattle after superovulation with Folltropin or Pluset

Two commercial FSH products were compared in a retrospective study on 3990 commercial superovulations and embryo recoveries in dairy heifers and cows. In addition, the 56-day nonreturn rate of 19,400 embryos produced with these two preparations was analyzed. Embryo collections were performed during a 16-year period from donors of Holstein and Ayrshire breeds. Folltropin (Vetoquinol S.A., Lure cedex, France) group (Group F) consisted of 2592 superovulations, of which 80% were performed on heifers and 20% on cows, and Pluset (Laboratorios Calier, S.A., Barcelona, Spain) group (Group P) of 1398 treatments, of which 66% and 34% were on heifers and cows, respectively. Total number of recovered structures, number of transferable embryos, and the proportion of unfertilized ova (UFO) and degenerated embryos were analyzed. Distribution of embryos into quality grades (1–3) and developmental stages (4–9) according to the IETS classification guidelines and means for each collection were evaluated. The proportion of low-responders having fewer than five corpora lutea and yielding fewer than five embryos or ova was investigated for each treatment. Group P yielded 1.1 recovered structures more than Group F (P < 0.001). Consequently, however, the number of transferable embryos did not differ among the groups, being 7.0 and 7.1 in Groups F and P, respectively. Instead, there was an increase in the number of UFO from 2.0 in Group F to 3.0 in Group P (P < 0.001). The quality of embryos and the developmental stages were similar between the groups and there was no difference in the proportion of low-responding donors in Group F and Group P. Also, there was no difference in the nonreturn rate after transfer of embryos originating from donors superovulated with Folltropin or Pluset. It was concluded that equal numbers of transferable embryos and pregnancies can be achieved with Folltropin and Pluset.

Effect of supplementation of hCG or GnRH on ovulation and subsequentembryo production of eCG superovulated goats

The aim of this study was to enhance the ovulation and subsequent embryo production of equine chorionic gonadotrophin (eCG) superovulated goats by supplementing with human chorionic gonadotrophin (hCG) or gonadotrophin releasing hormone (GnRH). Thirty crossbred donor does were oestrus synchronized using controlled internal drug release (CIDR) device and superovulated using 1500 IU eCG. Then all the donor does were randomly divided into 3 groups. Group 1 were administered 500 IU hCG each at 2 successive days starting from 24 hours after CIDR removal; Group 2 received a single dosage of 20 μg GnRH and Group 3 was control. Ovulation and embryo production responses were evaluated during laparotomy session on Day 7 after CIDR removal. Does in Group 1 showed higher number of CL (10.90) than the Group 2 (1.90) or Group 3 (0.90). The number of ovarian stimulation and number of anovulatory follicles had no significant (P&gt;0.05) differences among the treatment groups. Average number of structure recovered was significantly (P&lt;0.05) higher in Group 1 (3.10) than Group 2 (0.70) or Group 3 (0.00). Moreover, average number of embryo production was higher in Group 1 (0.90) than Group 2 (0.50) and Group 3 (0.00), but no significant difference was observed between Group 1 and Group 2. Results indicated that using hCG hormone would be an effective means for increasing ovulation in eCG superovulated crossbred goats, although it was unable to enhance the embryo number due to increase number of unfertilised ovum.

Identification and structural characterization of two peroxisome proliferator activated receptors and their transcriptional changes at different developmental stages and after feeding with different fatty acids.

Peroxisome proliferator activated receptors beta1 (PPARβ1) and beta2 (PPARβ2) were investigated in loach (Misgurnus anguillicaudatus). The PPARβ1 and PPARβ2 were widely distributed in loach tissues. Multiple alignments of deduced amino acid sequences revealed homologous characteristics of the two subtypes of PPARβ with 88% identity. PPARβ1 was markedly expressed in the liver, about 100-fold higher than liver PPARβ2. The two subtypes in unfertilized ovum (UO) showed the highest transcriptions in early life stages, and there were great divergences in expression between unfertilized and fertilized stages. The regulation of PPARβ1 and PPARβ2 in response to dietary fatty acids was studied in liver of loach fed with diets containing fish oil (FO, rich in n-3 highly unsaturated fatty acid) or soybean oil (SO, rich in 18:2n-6) for 75days. Results showed that hepatic transcription of PPARβ1 in the SO group was higher than in the FO group. However, PPARβ2 expression was similar. The differences of molecular characterization, tissue expressions in early life stages, and transcriptional regulation by lipid resources indicated that PPARβ1 and PPARβ2 were functionally different. This is the first report of differential expression of PPARβ1 and PPARβ2 in various tissues and early life stages of loach are regulated by lipid resources. These results will stimulate further studies to better understand the functional characterization of PPARβ1 and PPARβ2.

233 ANTI-MÜLLERIAN HORMONE AS A PREDICTIVE ENDOCRINE MARKER FOR SUPEROVULATORY RESPONSE AND EMBRYO PRODUCTION IN BEEF CATTLE

A study investigated the use of anti-Müllerian hormone (AMH) and/or follicle counts as a predictor of subsequent superovulatory response and embryo production in 79 beef cows. Before initiation of superovulation, ultrasonography was used to scan the ovaries of each donor cow to record the number of 3 to 5 mm follicles present, and a blood sample was collected for measure of serum AMH. At the time of embryo collection, the ovaries of donor cows were palpated to estimate the number of corpora lutea (CL) present on each ovary. Recovered embryos were evaluated for stage of development and morphological quality. Across cows, serum AMH ranged from 0.013 to 0.898 ng mL–1, with a mean of 0.293 ng mL–1. The distribution of AMH concentrations was divided into quartiles (AMH Q1 through Q4, with Q1 the lowest and Q4 the highest, ng mL–1) for analysis (ANOVA). Number of donors/collections in AMH Q1 through Q4 was 26, 22, 24, and 24, respectively. Donor cows in AMH Q4 had a greater (P &lt; 0.001) number of 3- to 5-mm follicles at the start of superovulation than did donors in either Q1 or Q2. At embryo collection, cows in AMH Q3 and 4 had more (P &lt; 0.001) palpable CL than cows in AMH Q1. The mean number of embryos recovered from donor cows in AMH Q4 (20.1 ± 1.8) was greater (P &lt; 0.001) than those recovered from cows in either AMH Q1 (9.8 ± 1.8) or 2 (9.4 ± 1.9), but similar to that of AMH Q3 (15.5 ± 1.8). Percentages of recovered embryos classified as transferrable, degenerate, or unfertilized were similar (P ≥ 0.275) across AMH quartiles. Analysis indicated that AMH was positively correlated (P &lt; 0.001) with mean follicles (r = 0.458), CL (r = 0.452), and embryos recovered (r = 0.430). To determine if follicle counts at the start of superovulation are predictive of superovulatory response, the distribution of follicle counts was divided into quartiles (F Q1 through Q4, with Q1 the lowest and Q4 the highest) for analysis. Donor cows with higher follicle counts (F Q3 and 4) at the start of superovulation had more (P &lt; 0.001) palpable CL at embryo collection than donor cows in F Q1 or 2. More (P &lt; 0.001) embryos (20.0 ± 2.2 v. 9.6 ± 1.8 and 11.6 ± 1.6) were recovered from cows with the highest follicle counts (F Q4) compared with cows having lower (F Q1 and 2) follicle counts, respectively. The percentage of transferable embryos and unfertilized ova were similar (P ≥ 0.688) across follicle count quartiles. As was noted for AMH, mean number of follicles at the start of superovulation was positively correlated (P &lt; 0.001) with mean CL (r = 0.556) and mean embryos (r = 0.423) but not percentages of viable or degenerate embryos or unfertilized oocytes (P ≥ 0.153). Results confirm that relative AMH concentration was positively correlated with number of small antral follicles in the ovaries of cows and might be used to either predict superovulatory response or possibly adjust superovulatory regimen to improve superovulatory response. Further study is needed to determine the effectiveness of using either AMH concentration or follicle counts to adjust superovulatory regimens.

Comment on “International Stem Cell”: The EU Court of Justice Revisits the Patentability of Processes for the Production of Human Stem Cells – The Decision in International Stem Cell Corporation

The brocard ius excludendi omnes alios describes the essence of patentability. These four words define a very basic principle: each patent confers to the owner a crystal-clear right to exclude others from the utilisation of the invention. However, as we move from the general ground of patentability to the specific field of human stem cells (hSCs), this crystal starts to reveal blurry edges and opacities. The aim of this paper is to shed light on these opacities. The recent decision of the Court of Justice of the European Union (CJEU) in International Stem Cell Corporation v. Comptroller General of Patents, Designs and Trade Marks will be used as a means to interpret hSCs patentability. The CJEU returned, after its decision in Brustle, on the definition of a human embryo expunging any unfertilised human ovum whose division and further development have been stimulated by parthenogenesis.

Coleochaete and the origin of sporophytes.

• Zygotes of Coleochaete are provisioned by the maternal thallus before undergoing 3-5 rounds of division to produce 8-32 zoospores. An understanding of the selective forces favoring postzygotic divisions would be relevant not only to the interpretation of Coleochaete life history but also to the origin of a multicellular diploid phase in embryophytes.• Simple optimization models are developed of the number of zygotes per maternal thallus and number of zoospores per zygote.• Zygotic mitosis is favored once zygotic size exceeds a threshold, but natural selection usually promotes investment in additional zygotes before zygotes reach this threshold. Factors that favor production of fewer, larger zygotes include multiple paternity, low fecundity, and accessory costs of zygote production. Such factors can result in zygotes exceeding the size at which zygotic mitosis becomes profitable.• Coleochaete may possess large zygotes that undergo multiple fission because of accessory costs associated with matrotrophy, including costs of cortical cells and unfertilized oogonia. The unpredictability of fertilization on land is proposed to have increased accessory costs from unfertilized ova and, as a consequence, to have favored the production of larger zygotes that underwent postzygotic division to produce diploid sporophytes.