A study on superovulation using FSH and eCG in Awassi ewes

The present study was conducted to evaluate superovulatory treatments in Awassi ewes by eCG and FSH. High number of unovulated follicles (P < 0.05) was observed in ewes treated with eCG in non-breeding season. It could be concluded that using FSH to induce superovulation in Awassi ewes is better than eCG.     

FSH injections and intrauterine insemination in protocols for superovulation of ewes

In Exp. 1, five injections of crude FSH (20 mg total), starting 36 h before removal of progestin pessaries or injection of prostaglandin F2 alpha (PGF2 alpha), were found to be as effective for superovulation of ewes as seven injections starting 72 h before removal of pessaries. Seven injections caused superovulation (more than three ovulations) in 13 of 15 ewes with an average (all ewes) of 12 +/- 2.3 ovulations. Five injections caused superovulation in 17 of 19 ewes with an average of 11.5 +/- 1.6 ovulations after pessary-regulated estrus in 12 of 16 ewes with an average of 12.4 +/- 2.2 ovulations after PGF2 alpha injection. In Exp. 2, intrauterine artificial insemination (IUAI) of naturally mated ewes provided ovum fertilization rates of 75.3% with an embryo recovery rate of 39.4%, whereas mating alone (MATED) provided fertilization rates of 64.2% with an embryo recovery rate of 72.1%. However, subsequent replications of the MATED protocol (MATED2) resulted in a low fertilization rate (34.2%). In Exp. 3, surgical IUAI at 22 h after removal of pessaries resulted in a low rate of superovulation (5 of 12 ewes), presumably as the result of pentobarbital anesthesia. Conducting IUAI at 46 h after removal of pessaries resulted in a high rate of superovulation (14 of 16 ewes) but a low embryo recovery rate (30.7%). In two subsequent trials, IUAI at 40 h after removal of pessaries resulted in most ewes superovulating (33 of 35 in April/May and 16 of 17 in September) with acceptable embryo recovery (65.2% and 71.3%) and high fertilization rates (82.8% and 96.4%).(ABSTRACT TRUNCATED AT 250 WORDS)     

A preliminary study on the suitability of Cervidil to induce cervical dilation for artificial insemination in ewes

The aim of this study was to evaluate the efficacy of Cervidil^®, a prostaglandin E₂ (PgE₂)-releasing vaginal insert used for induction of cervical ripening and labour in women, to enhance the ease of transcervical artificial insemination (TCAI) in anestrous ewes (June). It was hypothesized that the use of Cervidil^® prior to AI would cause dilation of the cervix, and thus alleviate the difficulty associated with traversing the cervix for semen deposition in sheep. Cervidil^® was inserted 12 h before insemination in six Rideau-Arcott ewes; six ewes served as untreated controls.Semen was deposited into the uterus of all six treated ewes but TCAI was possible only in four of the six control ewes. It can be concluded that Cervidil^® facilitated transcervical semen deposition in anestrous ewes. The treatment with Cervidil^® has the promise of a technique to improve transcervical AI and to enable non-invasive embryo transfer procedures in sheep.     

Comparison of three superovulation protocols with or without GnRH treatment at the time of artificial insemination on ovarian response and embryo quality in Thai native heifers

Tropical Animal Health and Production - To optimize the superovulation protocol in Thai native cattle, the present research was designed to (1) compare three different protocols designed to induce...     

The effect of melatonin treatment on the ovarian response of ewes to the ram effect

To determine the ovarian response to the ram effect after treatment with melatonin, on 8 March, 71 Rasa Aragonesa ewes were randomly assigned to either the treatment group and given an 18mg melatonin implant or the untreated group. On 19 April (day 0), rams were introduced into the flock. Melatonin treatment produced a significantly higher percentage of cyclic ewes at ram introduction (P<0.05). Melatonin-treated ewes had their first oestrus after ram introduction significantly earlier than did untreated ewes (P<0.0001), and the groups differed in the distribution of their ovarian response. Most (80%) of the treated ewes exhibited a silent ovulation followed by a cycle of normal duration, whereas about half (52%) of the untreated ewes exhibited a silent ovulation, a short cycle, and another silent ovulation followed by a cycle of normal duration (P<0.05). At ram introduction, melatonin-treated ewes, cyclic and non-cyclic, had higher mean plasma progesterone concentrations than did untreated ewes. The proportion of ewes that mated within the first 17 days of the mating period was significantly higher among the treated ewes than in the untreated ewes (P<0.0001). Furthermore, at lambing, 39% of the melatonin-treated ewes lambed within the first 17 days of the lambing period, while none of the untreated ewes lambed in that period. The untreated group exhibited peaks in mating between days 18 and 21, and particularly, between days 22 and 25, when the majority of ewes lambed; peaks did not occur in the treatment group. Treated and untreated ewes did not differ significantly in fertility, litter size and fecundity. In conclusion, melatonin treatment modifies the ovarian response to the ram effect in ewes, which leads to modifications in mating patterns, and consequently, the lambing curve.     

Superovulatory response and embryo quality in Katahdin ewes treated with FSH or FSH plus eCG during non-breeding season

The aim of this study was to evaluate the effect of a co-treatment of follicle-stimulating hormone (FSH) plus equine chorionic gonadotrophin (eCG) on serum insulin and insulin-like growth factor 1 (IGF-1) concentrations, superovulatory response, ovulatory rate, and number and production of embryos in Katahdin breed ewes during the non-breeding season. Twenty Katahdin ewes were synchronized with progestagens (CIDR) and assigned to two superovulation treatments (n = 10): (1): ewes treated with 200 mg ewe⁻¹ of FSH from day 5 to 8 after CIDR insertion at decreasing doses every 12 h (FSH group) and (2) ewes treated as FSH group plus 300 IU of eCG on day 5 after CIDR insertion (FSH + eCG group). Estrous behavior was monitored and direct mating was performed. On days − 7 (CIDR insertion), 0 (CIDR withdrawal), and 7 (embryo recovery), blood samples were collected to determine serum hormone concentrations. Co-treatment with eCG (FSH group) did not affect (P > 0.05) serum hormone levels. Superovulation response, ovulation rate, recovery rate, fertilization, and number of embryos were also similar (P > 0.05) between treatments. Compared with FSH group, FSH + eCG ewes had lower (P < 0.05) number of transferable embryos and higher (P < 0.05) number of oocyte and a tendency to increase the number of degenerated embryos (P = 0.07). Overall results suggest that the administration of eCG is not beneficial either to improve the ovulatory response or the amount of transferable embryos in Katahdin ewes superovulated with a protocol using progesterone and FSH at decreasing doses.

     

Preliminary study of ovarian activity in fillies treated with a GnRH vaccine

Objective To investigate the effects of two doses (200 and 400 mg) of a water-soluble gonadotrophin-releasing hormone vaccine on the ovarian activity of 2-year-old fillies.
Design A controlled vaccination dose rate experiment.
Animals Six 2-year-old Australian Stock Horse fillies were randomly allocated to three treatment groups: unvaccinated controls, those receiving 200 mg of the vaccine and those receiving 400 mg of the vaccine.
Results Ovarian activity of the treated fillies was suppressed at the peak of the breeding season while that of the untreated controls continued normally. The control fillies displayed oestrous activity and behaviour. Suppression of ovarian activity occurred for 25 and 30 weeks in the 200 and 400 mg groups, respectively. These differences were not significant. Ovarian activity ceased 2 to 3 weeks after primary vaccination. Antibody titres were low (330) until after the booster immunisation when they rapidly peaked at 22,000 and 28,000 in the 200 mg and 400 mg groups, respectively. Plasma progesterone concentrations of the treated fillies remained below 3.18 nmol/L while GnRH was suppressed. The vaccine had no significant effect on plasma androstenedione concentrations. Recovery from the effect of the vaccine was associated with development of ovarian follicles, normal oestrous behaviour and ovulation. Three of the four treated fillies and one of the controls conceived during the next breeding season and foaled normally. All the treated fillies conceived and produced normal foals in the following two breeding seasons.
Conclusion Both dose rates suppressed ovarian function and prevented oestrous behaviour. These effects were reversible and the subsequent fertility of the vaccinated fillies was normal.     

High‐milking sheep have a lower ovulation rate and tend to yield fewer embryos in response to superovulation and intrauterine artificial insemination

Antagonistic relationship between milk yield and reproduction is reported in several livestock species. This study aimed to investigate whether genetic merit for milk production in dairy sheep affects responses to superovulation, embryo yield and quality. A total of 21 cross‐bred Sarda x Lacaune ewes homogeneous for age, parity and stage of lactation were included. The ewes were stratified as high‐producing or low‐producing based on their genetic merit for milk production estimated by a pentatrait repeatability animal model. Oestrus was synchronized using an intravaginal progesterone pessary inserted on Day 0 and removed on Day 14. Superovulatory treatment consisted of 350 I.U. of porcine FSH administered in eight decreasing intramuscular doses every 12 hr with a total dose of 10 ml of solution starting 12 days after insertion of sponges. Laparoscopic artificial insemination (AI) was performed 48 hr after pessary removal. Surgical embryo recovery was performed at Day 8 after pessary removal. Correlation between breeding value for milk production and the number of corpora lutea (CL) was significantly different from zero (?0.49). High‐producing ewes had a lower number of CL than low‐producing counterparts (7.6 ± 2.50 vs 12.1 ± 5.16 respectively; p < .02). Furthermore, there was a tendency for high‐producing ewes to yield fewer embryos than low‐producing females (5.3 ± 3.46 vs 9.18 ± 5.11; p = .09). No differences were observed between ewes in both genetic groups with regard to the number of embryos of grades 1, 2 and 3. To our knowledge, this is the first report highlighting an antagonism between genetic merit for milk production and the ability to produce embryos in sheep. These results deserve to be considered in sheep breeding programmes.     

The effect of hormonal preparations on superovulation and egg transport in ewes

The relation between the prevalence of contagious mastitis with the finding of Streptococcus agalactiae and Staphylococcus aureus and the number of somatic cells in the tank samples of milk was studied on the basis of a clinical and bacteriological examination of 11 129 dairy cows on four large farms, with a capacity of 600 to 1240 animals. A statistically significant correlation was demonstrated between the analyzed characteristics (r = 0.621). The relation was demonstrated at a prevalence up to 25.3%. The number of somatic cells provides a marked indication of prevalence only from 10%. We failed to demonstrate a statistically significant correlation between the prevalence of mastitis with a cultivation finding significant correlation between the prevalence of mastitis with a cultivation finding of Staphylococcus aureus and the number of somatic cells in tank samples of milk (r = 0.054). The results suggest that on large farms it will not be possible to use only the number of somatic cells in tank milk samples as a signal of diagnostic improvement and preventive mastitis-control measures, since a 10% prevalence of these diseases usually excludes the possibility of herd sanitation.     

Effect of reproductive methods and GnRH administration on long-term protocol in Santa Ines ewes

This study aimed to determine whether reproductive performance of ewes submitted to laparoscopic timed artificial insemination (TAI) would be similar to ante meridiem (AM)/post meridiem (PM) rule and assisted natural mating (NM), and whether GnRH may enhance the pregnancy rate in TAI. In experiment I, 191 non-lactating ewes were synchronized, then TAI was performed either 48 h after progesterone (P4) removal (TAI-48 h) or 12 h after estrus detection (AM/PM); moreover, some ewes were submitted to NM (NM) as control treatment. In experiment II, 247 non-lactating ewes were allocated in five treatments, a control (no-GnRH on protocol) and four treatments arranged in a factorial design 2 × 2. The factors were time and dose of GnRH: ewes that received either 10 μg (TAI-10 μg-36 h) or 25 μg of GnRH (TAI-25 μg-36 h) 36 h after P4 removal and ewes that received either 10 μg (TAI-10 μg-48 h) or 25 μg of GnRH (TAI-25 μg-48 h) at time of insemination, 48 h after P4 removal. In experiment I, pregnancy rate in TAI-48 h was lower (P = 0.03) than AM/PM and NM. Moreover, the probability of pregnancy in TAI-48 h was higher (P = 0.06) in ewes detected in estrus early. In experiment II, the use of GnRH in TAI protocols increased (P < 0.01) pregnancy rate at synchronization, and TAI-25 μ-48 h and TAI-10 μg-36 h treatments increased (P = 0.02) pregnancy rate compered to TAI-10 μg-48 h. We conclude that TAI decreased pregnancy rate compered to NM and AM/PM, which may be improved by GnRH use in TAI to synchronize ovulation.

     

The effect of high calcium intake by pregnant Awassi ewes at late gestation on minerals status and performance of ewes and newborn lambs

Calcium (Ca) requirement increase rapidly by pregnant ewes at late gestation as a result of high fetal growth. Furthermore, Ca deficiency results in poor growth and bone development in growing newborn lambs. The aim of this study was to evaluate the effect of calcium supplementation (Calcium carbonate–CC) and vitamin AD₃E injection (75,000 IU/shot) to pregnant Awassi ewes at late gestation (− 45 days prepartum) on their mineral levels and performance of their newborn lambs. Forty pregnant Awassi ewes at late gestation (65 ± 6.1 kg) were selected before parturition (− 45 days; using hormonal sponges for estrus synchronization) and randomly divided to 4 dietary treatments. The treatments were: group 1 Control (1.4% CC); group 2 (2.4% CC); group 3 (1.4% CC + 2 shots vitamin AD3E at − 45 and − 21 days prepartum); group 4 (2.4% CC + 2 shots vitamin AD3E at − 45 and − 21 days prepartum). Blood samples were collected from ewes (− 45, 0 days prepartum: 30 and 60 days postpartum) and lambs (0, 30 and 60 days old). Average milk yield and samples were collected. Body weights of lambs were taken at birth, 30 and 60 days old. Blood serum and milk samples were prepared and analyzed by AAS for Ca, Mg, Co, Mn and Cu concentrations.Calcium concentrations in bloods serum of ewes, 0 and 60 days postpartum, and lambs at weaning (60 days old) from group 4 were significantly (P < 0.05) higher compared to other groups. No significant effect (P > 0.05) of treatment on Co, Mg and Cu in blood serum of ewes and lambs except for Cu concentration in ewes. Moreover, a significant increase (P < 0.01) in milk inorganic% (30 and 60 days samples) in ewes from group 2 when compared with other groups. The average milk yield (AMY) of ewes from group 2 and 4 at late stage of lactation (45–60 days) were significantly higher (P < 0.05) compared to groups 1 and 3. In addition, Ca% in colostrums samples from ewes in groups 2 and 4 were significantly higher (P < 0.01). The average daily gain (ADG) and total body gain for lambs up to weaning (60 days old) were significantly higher (P < 0.05) in groups 1 and 2 compared with the other groups.In conclusion, increasing level of Ca intake in feed of Awassi ewes at late gestation, with or without vitamin AD₃E injections, improve the calcium concentration in colostrums and blood serum of ewes and their lambs without negative effect in their health and performance.     

高寒放牧条件下牦牛超排试验

在高寒放牧条件下 ,对 12头祁连山型母牦牛进行了选择不同超排方法和FSH剂量及使用中科院动物所FSH、LH ,国产PGF2α、HCG ,新西兰FSH的超排试验 ,供试母牦牛在发情 11～ 12天开始注射FSH。结果表明 :A组 :FSH 8 4mg ,母牦牛与正常发育相似 ,冲出 2枚发育正常囊胚 ,1头放弃 ;B组 :FSH 8 6mg ,利用PVP包埋 1次注射 ,母牦牛卵巢处于静止状况 ,插管 1头 ,没有冲出胚胎 ,超排处理无结果。说明牦牛用FSH 8 4～ 8 6mg剂量偏小 ,对超排无效果 ;C组 :FSH 8 8mg ,分FSH +LH( 2 0 0IU)、FSH +HCG( 10 0 0IU)、卵巢黄体数 10± 4,2头牦牛冲卵 ,获 14枚胚胎 ,有 1头牦牛冲出 10枚胚胎 ,其中有 2枚发育到 16细胞 ,1枚发育到 3 2细胞 ,说明FSH +HCG(LH)有协同作用 ,促进卵巢上卵母细胞成熟和排卵 ;D组 :FSH 15mg(新西兰产 ) 1头牦牛冲出 3枚发育正常早期囊胚。     

Changes in the concentration of mRNAs for the inhibin subunits in ovarian follicles after administration of gonadotropins to progestin treated ewes.

RNA was extracted from single or small groups of ovine ovarian follicles after treatment of ewes with FSH and/or LH. The content of mRNA for the alpha-inhibin and beta A-inhibin subunits was analyzed by hybridization with specific cDNA probes. All ewes were treated with progestin vaginal pessaries to suppress spontaneous preovulatory follicle maturation and ewes were given three intramuscular injections of gonadotropins at 8-hr intervals starting 24 hr prior to collection of ovaries. In experiment I, both Schering-FSH and NIDDK-oFSH-17 (oFSH) significantly increased alpha- and beta A-inhibin mRNA per ewe in 2-5 mm follicles and tended to increase alpha- and beta A-inhibin mRNA in large (greater than 5 mm) follicles. In experiment II, oFSH and NIDDK-oLH-25 (oLH) were administered in a 2X2 factorial arrangement. Separate administration of oFSH or oLH increased (P less than .05) the alpha-inhibin mRNA concentration in large follicles. alpha-inhibin mRNA concentration in 4-5 mm follicles was also increased by oFSH but was decreased by oLH. Concomitant treatment with oFSH and oLH did not change alpha-inhibin mRNA concentrations from those measured in oFSH treated ewes. In experiment II, beta A mRNA concentrations followed a pattern similar to that of alpha A mRNA, but the differences were not statistically significant. We conclude that, in the ewe, exogenous FSH increases the concentration of inhibin mRNA in the whole follicle. The ability of exogenous oLH to alter expression of the inhibin subunit genes may depend upon the stage of follicle maturation.     

Basal and GnRH induced secretion of FSH and LH in ovariectomized ewes treated with bovine follicular fluid

Ovariectomized (OVX) ewes were injected with 5 ml of either bovine serum, charcoalextracted bovine follicular fluid (FF), or whole bovine FF. Five hours after this pretreatment, ewes on each pretreatment were injected with either 0, 1, or 5 μg of GnRH. Ewes that were pretreated with either type of FF had decreased concentrations of FSH regardless of dose of GnRH when compared to ewes pretreated with bovine serum. There was no effect of charcoal extraction. There were no differences among the pretreatment groups in LH response to GnRH. In a second experiment, OVX ewes were pretreated (4 ml) with either bovine serum or bovine FF 5 hr prior to GnRH or with bovine FF 42, 30 and 18 hr prior to GnRH. Ewes were injected with either 0 or 5 μg of GnRH. Pretreatment with FF for 5 or 42 hr prior to GnRH resulted in significantly decreased concentrations of FSH both at the time of GnRH treatment and during the following 2 hr. Concentrations of LH did not differ among pretreatment groups. In a third experiment, OVX ewes were pretreated with either bovine serum or bovine FF 30, 18 and 5 hr prior to GnRH. Ewes were injected with either 0, 5 or 50 μg of GnRH. Pretreatment with FF resulted in decreased concentrations of FSH both at the time of GnRH treatment and during the following 2 hr. Concentrations of LH were also decreased at the time of GnRH treatment.     

Endocrine and ovarian changes in response to the ram effect in medroxyprogesterone acetate-primed Corriedale ewes during the breeding and nonbreeding season

Two experiments were performed to determine the endocrine and ovarian changes in medroxyprogesterone acetate (MAP)-primed ewes after ram introduction. Experiment 1 was performed during the mid-breeding season with 71 ewes primed with an intravaginal MAP sponge for 12 days. While the control (C) ewes (n = 35) were in permanent contact with rams, the ram effect (RE) ewes (n = 36) were isolated for 34 days prior to contact with rams. At sponge withdrawal, all ewes were joined with eight sexually experienced marking Corriedale rams and estrus was recorded over the next 4 days. The ovaries were observed by laparoscopy 4-6 days after estrus. Four weeks later, pregnancy was determined by transrectal ultrasonography. In eight ewes from each group, ovaries were ultrasonographically scanned; FSH, LH, and estradiol-17beta were measured every 12 hours until ovulation or 96 hours after estrus. The response to the rams was not affected by the fact that ewes had been kept or not in close contact with males before teasing. No differences were found in FSH, LH, estradiol-17beta concentrations, growth of the ovulatory follicle, onset of estrus, ovulation rate, or pregnancy rate. Experiment 2 was performed with 14 ewes during the nonbreeding season. Ewes were isolated from rams for 1 month, and received a 6-day MAP priming. Ovaries were ultrasonographically scanned every 12 hours, and FSH, LH, estradiol-17beta, and progesterone were measured. Ewes that ovulated and came into estrus had higher FSH and estradiol-17beta levels before introduction of the rams than did ewes that had a silent ovulation. The endocrine pattern of the induced follicular phase of ewes that came into estrus was more similar to a normal follicular phase, than in ewes that had a silent ovulation. The follicle that finally ovulated tended to emerge earlier and in a more synchronized fashion in those ewes that did come into estrus. All ewes that ovulated had an LH surge and reached higher maximum FSH levels than ewes that did not ovulate, none of which had an LH surge. We conclude that (a) the effect of ram introduction in cyclic ewes treated with MAP may vary depending on the time of the breeding season at which teasing is performed; (b) patterns of FSH, and estradiol-17beta concentrations, as indicators of activity of the reproductive axis, may be used to classify depth of anestrus; and (c) the endocrine pattern of the induced follicular phase, which is related to the depth of anestrus, may be reflected in the behavioral responses to MAP priming and the ram effect.     

Effect of season and photoperiod on the time of first postpartum ovulation in Awassi ewes

Seasonal differences in the resumption of postpartum ovarian activity, milk production and periparturient metabolic status were investigated in lactating non-suckling dairy Awassi sheep in two consecutive experiments. In Experiment 1, autumn-lambing (AL, n = 27) and spring-lambing (SL, n = 37) ewes were investigated. Ovarian activity was monitored by means of individual progesterone (P4) profiles from day 5 to day 100 post partum. Most of the AL dams (89%) ovulated till day 35 after parturition and became cyclic thereafter. Incidence of persistent corpus luteum (CLP) and short luteal phases (sCL) was frequent (18% and 29%, respectively) among non-conceiving dams. In contrast, only 24% of the SL ewes ovulated before day 35. P4 levels during the luteal phase were lower in cyclic animals, and the cycle was longer in SL than in AL animals. No CLP or sCL was detected in the spring-lambing group, and 61% of SL ewes remained acyclic till the end of the trial. Lactation length was significantly longer in SL dams than in AL ewes (P = 0.008). According to the plasma metabolites (BHB, NEFA) and metabolic hormones (insulin, IGF-I, thyroxine) examined, negative energy balance did not appear in any of the animals. However, seasonal differences were seen in IGF-I and thyroxine levels, which were higher in the SL dams. In Experiment 2, influence of additional lighting was studied in autumn-lambing ewes. The long-day photoperiod (LD, n = 23) group was exposed to artificial light from sunset till midnight (approx. 16 h light/8 h dark) from some weeks before the expected date of delivery in mid-September until the end of December. The control group (n = 25) experienced only natural daylength. The first postpartum ovulation tended to occur later in the LD animals than in the controls (P = 0.047). The lactation of the LD group tended to be longer (P = 0.061). NEFA, BHB, insulin, IGF-I and thyroxine levels did not differ between the groups. Conclusions: (i) The ovarian function of the Awassi population is seasonal under temperate continental climate conditions. (ii) The first postpartum ovulation of non-suckling, autumn-lambing dams may occur very early, even before the completion of uterine involution. (iii) Additional artificial lighting may delay the time of first postpartum ovulation in AL ewes. (iv) Postpartum negative energy balance is unlikely to occur in dairy Awassi ewes even in high-producing intensive systems.