Preovulatory Follicle Size Research Articles

177 EFFECTS OF MANIPULATION OF DOMINANT FOLLICLE GROWTH ON SIZE AND FUNCTION OF CORPUS LUTEUM IN BEEF CATTLE

Recent evidence indicates that the progesterone (P4) secretion by corpus luteum (CL) during early diestrus is affected by the size of ovulatory follicle and has a significant impact on embryo development and conception rates. Therefore, strategies to promote the growth of the dominant follicle and/or to stimulate the early development of the CL to increase P4 secretion become an alternative to improve conception rates in the beef cattle industry. Our aim was to study the effect of manipulations of the follicle growth on the diameter of the preovulatory follicle (POF) and subsequent size and function of the CL. Cyclic and non-lactating Nelore cows, pre-synchronized by 2 injections of prostaglandin F2α (PGF) 14 d apart, were manipulated to ovulate large or small follicles according to 3 experiments. In Experiment 1 (Exp. 1; n = 23), animals received a second-use intravaginal P4-releasing device along with an injection of oestradiol benzoate on Day –10 (Day 0 = GnRH injection). Cows were split to receive (large follicle group; LF) or not (small follicle group; SF) a PGF injection on Day –10. Progesterone devices were removed on Day –2.5 in the LF group and on Day –1.5 in the SF group. The PGF was injected at the removal of the P4 device. In Experiment 2 (Exp. 2; n = 38), cows in the LF group had the P4 device removed on Day –2.25 or Day –2, whereas in Experiment 3 (Exp. 3; n = 23), the device (first-use) was removed on Day –1.75 in the LF group and on Day –1.25 in the SF group; the other manipulations were similar to Exp. 1. Data analyses were done only on cows that had a functional CL on Day –10 (P4 > 1 ng mL–1) and that ovulated within 24 and 48 h post-GnRH (Exp. 1, n = 14; Exp. 2, n = 14; Exp. 3, n = 12). The three experiments were successful in inducing POF with different sizes, as indicated by the greater diameter of the POF in the LF group compared with SF in Exp. 1 (12.9 ± 0.5 mm v. 10.7 ± 0.6 mm; P < 0.03), Exp. 2 (14.1 ± 0.6 mm v. 11.7 ± 0.4 mm; P < 0.006), and Exp. 3 (13.8 ± 0.6 mm v. 11.7 ± 0.8 mm; P < 0.06). To evaluate the effect of POF size on size and function of the CL, a factorial analysis was performed by SAS software to test the effect of group, day, and their interaction. For CL volume, an effect of group was detected in Exp. 1 (P < 0.02) and in Exp. 3 (P < 0.06), but not in Exp. 2. The group effect represented greater average CL volume from Day 3 to Day 7 in LF (2.42 ± 0.27 and 2.5 ± 0.39 cm3) than in the SF group (1.39 ± 0.18 and 1.2 ± 0.15 cm3) for Exp. 1 and 3, respectively. For P4 concentrations, a group effect was detected only in Exp. 3 (P < 0.007), as indicated by greater average P4 concentrations from Day 3 to Day 7 in LF (2.31 ± 0.31 ng mL–1) than in the SF group (1.37 ± 0.19 ng mL–1). A day effect was detected in all experiments (P < 0.0001), as indicated by a progressive increase of CL volume and P4 concentrations from Day 3 to Day 7. Manipulation of follicle growth performed in Exp. 3 was the most efficient to modify the function and size of the CL. In conclusion, control of POF size by manipulation of P4 concentrations during growth of the dominant follicle alters the size and function of CL postovulation. CNPq, FAPESP, Ourofino, and PUSP-P.

Effects of feeding regimen, fiber inclusion, and crude protein content of the diet on performance and egg quality and hatchability of eggs of broiler breeder hens

A 12-wk experiment was conducted to study the effects of feeding regimen, inclusion of a fiber source, and CP content of the diet on performance of broiler breeder hens. In total, 360 hens and 60 males, 43 wk of age, were assigned to 60 floor pens (6 hens and 1 male each). There were 12 treatments arranged factorially with 2 feeding regimens [restricted (R) and liberal feeding (close to ad libitum consumption; LIB)], 3 sources of fiber (0, 3% inulin, and 3% cellulose), and 2 levels of CP (14.5 and 17.4%). No interactions among main effects were observed for any of the traits studied, and therefore, only main effects are presented. Body weight, liver weight, and abdominal fat weight were higher (P < 0.001) for the LIB than for the R-fed hens. However, egg production (P < 0.001), fertility index (P < 0.05), and percentage of hatch (P < 0.01) were lower for LIB than for R hens. The weights of ovaries (P < 0.05) and the size of the first preovulatory follicle (P < 0.05) were higher for the LIB than for the R hens. Also, egg yolk, egg weight, and BW of the hatching chicks were higher (P < 0.001) for the LIB hens. The inclusion of a fiber source in the diet decreased (P < 0.05) feed intake, BW gain, absolute liver and abdominal fat weight, and egg yolk weight, with effects being more pronounced (P < 0.05) with cellulose than with inulin. Hens fed additional fiber produced more (P < 0.05) eggs that were more fertile (P < 0.05) than control hens. Crude protein content of the diet did not affect hen performance but reduced (P < 0.01) the relative weight of the liver, ovary, and abdominal fat. It is concluded that the inclusion of inulin or cellulose in the diet improved hen performance and that an increase in dietary CP reduced obesity in broiler breeder hens.

Comparison of Different Treatments for Oestrous Induction in Seasonally Anovulatory Mares

The aim of this study was to evaluate the effects of different treatments for induction and synchronization of oestrus and ovulation in seasonally anovulatory mares. Fifteen mares formed the control group (C), while 26 mares were randomly assigned to three treatment groups. Group T1 (n=11) were treated with oral altrenogest (0.044mg/kg; Regumate(®) ) during 11days. Group T2 (n=7) was intravaginally treated with 1.38g of progesterone (CIDR(®) ) for 11days. In group T3 (n=8), mares were also treated with CIDR(®) , but only for 8days. All mares received PGF2α 1day after finishing the treatment. Sonographic evaluation of follicles, pre-ovulatory follicle size and ovulation time was recorded. Progesterone and leptin levels were analysed. Results show that pre-ovulatory follicles were developed after the treatment in 88.5% of mares. However, the pre-ovulatory follicle growth was dispersal, and sometimes it was detected when treatment was not finished. While in mares treated with intravaginal device, the follicle was soon detected (1.5±1.2days and 2.3±2.0days in T2 and T3 groups, respectively), in T1 group, the pre-ovulatory follicle was detected slightly later (3.9±1.6days). The interval from the end of treatment to ovulation did not show significant differences between groups (T1=13.1±2.5days; T2=11.0±3.6days; T3=13.8±4.3days). The pregnancy rate was 47.4%, similar to the rate observed in group C (46.7%; p>0.05). Initial leptin concentrations were significantly higher in mares, which restart their ovarian activity after treatments, suggesting a role in the reproduction mechanisms in mares. It could be concluded that the used treatments may be effective for oestrous induction in mares during the late phase of the seasonally anovulatory period. Furthermore, they cannot synchronize oestrus, and then, it is necessary to know the reproductive status of mares when these treatments are used for oestrous synchronization.

Ovarian characteristics, serum concentrations of progesterone and estradiol, and fertility in lactating dairy cows in response to equine chorionic gonadotropin

We hypothesized that administration of eCG during the proestrous maturation phase of the preovulatory ovarian follicle would increase follicle size and serum estradiol concentrations. Our objectives were to evaluate the effects of eCG administration on preovulatory ovarian follicle size and growth rate, serum concentrations of estradiol and progesterone, estrual activity, posttreatment luteal activity, and pregnancy per AI. Lactating Holstein cows milked thrice daily were enrolled in a Presynch-Ovsynch timed AI program before the first AI. Cows (N = 128) in a single herd were given two doses of prostaglandin F2α (PGF2α) 14 days apart (Presynch), with the second dose 11 days before the onset of an Ovsynch protocol (treatment with GnRH 7 days before and 56 hours after PGF2α, with AI 16 to 18 hours after the second GnRH treatment). Cows were assigned randomly to receive either saline or 400 IU eCG concurrent with PGF2α treatment of the Ovsynch protocol (Day 0). Serum concentrations of progesterone and estradiol were assessed to determine if eCG would increase estrual activity, improve ovulatory response to GnRH, and enhance postovulatory luteal function. Compared with controls, treatment with eCG did not increase diameter or growth rate of the largest follicle during 48 hours after eCG, but tended (P = 0.09) to increase growth rate of the second largest follicle. Serum estradiol concentrations and estrual activity were not altered by eCG. Serum progesterone concentrations did not differ between treatments on Days 0, 2, 4, 9, or 16 after eCG treatment. Number of CL per cow on Days 9 and 16 after administration of eCG did not differ between treatments, but total luteal tissue volume tended (P = 0.06) to be greater on Day 16 for eCG-treated cows compared with controls. Pregnancy per AI at first service was similar between eCG (35.3%) and control cows (39.0%). We concluded that eCG treatment administered 3 days before insemination at the dose of 400 IU failed to increase follicle growth size and growth rate 48 hours after treatment, and did not enhance pregnancy outcomes in dairy cattle programmed for AI at first service.

Growth of the dominant follicle and endometrial folding after administration of hCG in mares during oestrus

The purpose of the trial was evaluation of follicular growth and endometrial folding in mares after human chorionic gonadotropin (hCG) treatment in comparison with untreated mares during oestrus. In addition, the influence of follicle size at the time of hCG treatment on these parameters was evaluated. HCG (3000 IU) was administered intravenously in 17 mares bearing dominant follicles 35&amp;ndash;40 mm in diameter (Group A) and in 13 mares with larger follicles (Group B). Ten mares with follicles &amp;ge; 35 mm were untreated (Group C). Ultrasonographical examination of the mares continued in 6 h intervals until ovulation. Growth of the dominant follicle was faster in Group A than in Groups B and C (1.3 vs. 0.3 and 0.7 mm/6h, P &amp;lt; 0.05) but diameters of the preovulatory follicles were similar &amp;ndash; 44, 48 and 44 mm in Groups A, B and C, respectively. Similarly, reduction of endometrial folding (on a three point scale) during observation was higher in Group A than in B and C (2.1 vs. 1.2 and 1.8, A : B P &amp;lt; 0.05) but endometrial folding values in the term before ovulation were not different (0.6, 0.9 and 0.6 in Groups A, B and C). A positive correlation between the speed of follicular growth and reduction of endometrial folding was found (r&lt;sub&gt;s&lt;/sub&gt; &amp;ndash; 0.479, P = 0.003). Irregularity in follicle shape (the difference between the longest axis and its perpendicular axis) at the beginning of observation (3.3, 4.0 and 3.2 mm) was lower than before ovulation (7.4, 10.4 and 9.2 mm) in all groups (P &amp;lt; 0.01). The interval from the beginning of observation until ovulation was significantly shorter in Groups A and B versus C (37 and 31 vs. 103 h, P &amp;lt; 0.01). The results show that growth of dominant follicles after hCG is influenced by the size of the follicles at the time of treatment and correlates with reduction in endometrial folding as well as irregularity of follicle shape. Nevertheless, hCG treatment does not influence the size and shape of preovulatory follicles or endometrial folding immediately before ovulation. &amp;nbsp;

Effects of synchronization treatments on ovarian follicular dynamics, corpus luteum growth, and circulating steroid hormone concentrations in lactating dairy cows

Lactating dairy cows (n=57) ≥45 d postpartum at first service were enrolled in a randomized complete block design study to evaluate treatments to synchronize estrus and ovulation. At 10 d before artificial insemination (AI), animals were randomly assigned to 1 of 3 treatments: (1) d −10 GnRH (GnRH1; 10μg of buserelin, i.m.) and controlled internal drug release insert [CIDR, 1.38g of progesterone (P4)]; d −3 PGF2α (PGF; 25mg of dinoprost, i.m.); d −2 CIDR out; and AI at observed estrus (CIDR_OBS); (2) same as CIDR_OBS, but GnRH (GnRH2) 36h after CIDR out and timed AI (TAI) 18h later (CIDR_TAI); or (3) same as CIDR_TAI, but no CIDR (Ovsynch). Transrectal ultrasound was used to assess follicle size before ovulation and on d 4, 8, and 15 after the presumptive day of estrus (d 0) to measure the corpus luteum (CL). Blood samples were collected to determine concentrations of estradiol (E2; d −10, −9, −3, −2, −1, and 0) and P4 (d −10, −9, −2, −1, 0, 1, 4, 6, 8, 11, and 15). No treatment differences were observed in either circulating concentrations of P4 or the ovulatory response to GnRH1 at the onset of synchronization treatments. Circulating concentrations of P4 were greater for CIDR_OBS and CIDR_TAI compared with Ovsynch at 24h after CIDR insertion (5.34 and 4.98 vs. 1.75 ng/mL) and immediately before CIDR removal (1.65 and 1.48 vs. 0.40 ng/mL). Peak circulating concentrations of E2 were greater for CIDR_OBS compared with Ovsynch (3.85 vs. 2.39pg/mL), but CIDR_TAI (2.82pg/mL) did not differ from either CIDR_OBS or Ovsynch. The interval from PGF injection to peak circulating E2 did not differ between CIDR_TAI and Ovsynch (52.1 vs. 49.8h). Both CIDR_TAI and Ovsynch, however, had shorter intervals from PGF injection to peak circulating E2 concentrations compared with CIDR_OBS (67.8h). The diameter of the dominant follicle before ovulation was greater for CIDR_OBS compared with Ovsynch (18.5 vs. 16.0mm) but CIDR_TAI (17.1mm) did not differ from either of the other treatments. The mean interval from PGF to ovulation was longer for CIDR_OBS (100.0h) compared with CIDR_TAI and Ovsynch (84.4 and 83.2h, respectively). Use of CIDR_OBS resulted in increased preovulatory follicle size and greater circulating concentrations of E2 due to a longer period of preovulatory follicle growth. Progesterone supplementation during synchronization and GnRH on the day before TAI affected ovulatory follicle size, and periovulatory circulating concentrations of P4 and E2. No differences, however, in postovulatory P4 or luteal volume profiles were observed.

Evaluation of ovarian toxicity of mono-(2-ethylhexyl) phthalate (MEHP) using cultured rat ovarian follicles

Mono-(2-ethylhexyl) phthalate (MEHP) is the most toxic metabolite of di-(2-ethylhexyl) phthalate (DEHP). It has been reported that DEHP causes abnormal reproductive development in women, and suppresses estradiol synthesis and ovulation in female rats with diminished size of preovulatory follicles. The present study was conducted to evaluate the ovarian toxicity of MEHP using cultured rat ovarian follicles. Secondary follicles were isolated from the ovaries of 14-day-old female rats and cultured for 48 hr with MEHP (0, 10, 30, and 100 µg/ml). At 0, 24, and 48 hr of MEHP treatment, follicular diameters were measured. After the culture, viability and apoptosis of follicles were assessed, and progesterone, androstenedione, testosterone, and estradiol levels in culture media were measured. At 100 µg/ml, suppression of follicular development was observed, which is associated with decreased viability of follicles and apoptosis of granulosa cells. At this concentration, progesterone level increased markedly, whereas androstenedione, testosterone, and estradiol levels decreased. At 10 and 30 µg/ml, follicular development was not suppressed, no apoptotic change was observed, and the levels of all measured steroid hormones tended to increase. The combined levels of all steroid hormones increased at all concentrations of MEHP, and the increase implies that MEHP activates the synthetic pathway from cholesterol to estradiol including de novo synthesis of cholesterol. However, the progesterone/androstenedione ratio increased extremely at 100 µg/ml, and the increase implies that MEHP inhibits the conversion of progesterone to androstenedione. In conclusion, MEHP induces ovarian toxicity via suppression of follicular development and abnormal steroid hormone synthesis in cultured rat ovarian follicles.

Influence of the Length of Progestagen Treatment and the Time of Oestradiol Benzoate Application on the Ovulatory Follicle Size and Ovulation Time in Anoestrous and Cyclic Beef Cows

Previous research from our laboratory in beef cattle suggests that the pre-ovulatory follicle size, maturity and subsequent susceptibility to gonadotropin are influenced by the length of progestagen treatment in artificial insemination programme in beef cows. To test this hypothesis, two experiments were conducted. In experiment 1, 35 anoestrous beef cows received an intravaginal sponge containing 200 mg of medroxyprogesterone acetate. The treatment lasted for 7 (n = 12), 8 (n = 11) or 9 (n = 12) days. Half of the animals in each group were injected with 0.7 mg of oestradiol benzoate (EB) at device removal (0 h) and the other half 24 h later. In experiment 2, 38 cycling beef cows were treated with the same protocols as in experiment 1. Ultrasound examinations were performed to determine the follicular diameter at device removal (dominant follicle), interval to ovulation and ovulatory follicle diameter. The dominant follicle of anoestrous cows with progestagen for 7 days (8.4 ± 1.6 mm) resulted smaller (p < 0.05) than the cows treated for 8 (10.5 ± 1.6 mm) and 9 days (10.6 ± 1.2 mm). However, regardless of the length of the treatments, ovulation time after device removal was longer (p < 0.05) when EB was injected 24 h after withdrawal than at 0 h in anoestrous cows (EB0 = 52.7 ± 4.0 h; EB24 = 70.8 ± 6.2 h) and in cyclic cows (EB0 = 50.0 ± 21.0 h; EB24 = 73.0 ± 20.0 h). In anoestrous cows, the treatment with progestagen for 9 days and EB at 24 h increased the diameter of the ovarian follicle (p = 0.033) but did not affect the diameter of the ovulatory follicle in cyclic cows. In conclusion, increasing the length of progestagen treatment for 8 or 9 days compared to 7 days increased the diameter of the dominant follicle, in anoestrous and cyclic beef cows. Oestradiol benzoate administered at device removal resulted in a shorter interval from device removal to ovulation compared with EB injection 24 h after the end of a progestagen treatment.

Factors affecting pre-ovulatory follicle diameter in the mare: the effect of mare age, season and presence of other ovulatory follicles (multiple ovulation)

The importance of elucidating factors affecting reproductive performance and efficiency is of paramount concern to the equine industry. Oocyte viability is known to be one of the determinants of reproductive success and evidence suggests that it may be linked to follicle size. The aims of this study were, therefore, to ascertain: i) the average diameter and range of pre-ovulatory follicles in Thoroughbred mares; ii) whether this is affected by either mare age, time within the breeding season, or the presence of multiple pre-ovulatory follicles (MO). One thousand, four hundred and ninety two Thoroughbred mares, aged 2–26 years, were examined with ultrasound to ascertain ovulation date to within 24h, and pre-ovulatory follicle(s) (F1) diameter. Mares were divided into groups according to age (7 groups, 2–4 yr, 5–7 yr, 8–10 yr, 11–13 yr, 14–16 yr, 17–19 yr, >19 yr), time within the season (16 half-month groups, from Feb 1 st to Sept 30 th), and pre-ovulatory follicles (single, {SO} or multiple {MO}). Overall average F1 diameter was 39.95 ± 4.84 mm (range 22–50 mm). Mare age had a significant (P < 0.001) negative effect on F1 diameter (largest F1 38.95 ± 5.61 mm, mares 2–4 yrs; smallest F1 33.30 ± 4.66 mm, mares >19 yrs) as did season (largest F1 44.20 ± 3.95 mm, Feb 1 st–14 th; smallest F1 33.74 ± 4.87 mm, Aug 15 th–31 st) and the presence of more than one pre-ovulatory follicle (MO F1 35.45 ± 4.53 mm; SO F1 37.44 ± 4.84 mm). In conclusion older mares, bred towards the end of the breeding season, especially if MO were present, were more likely to ovulate from smaller follicles. If, as suggested, small pre-ovulatory follicle size is associated with low oocyte viability, then this may account, at least in part, for the poor fertility rates characteristic of older MO mares, bred later in the season and so justify increased monitoring and careful reproductive management of such mares.

Post-weaning Altrenogest treatment in primiparous sows; the effect of duration and dosage on follicular development and consequences for early pregnancy

Our objective was to investigate follicle development in sows during and after different Altrenogest treatments post-weaning and relate this to subsequent ovulation rate and embryonic development. Primiparous UPB sows ( n = 47) were randomly assigned to (weaning = Day 0): control (no Altrenogest, n = 12), RU8-15 (15 mg of Altrenogest, Day-1 till Day 7, n = 12), RU8-20 (20 mg of Altrenogest, Day-1 till Day 7, n = 12) or RU15-15 (15 mg of Altrenogest, Day-1 till Day 14, n = 11). From weaning onwards, trans-abdominal ultrasound was performed daily. Sows were slaughtered on Day 4 or 5 after ovulation. Follicle size increased during Altrenogest treatment and reached a plateau around Day 6, regardless of dose (4.6 ± 1.5, 4.6 ± 1.6 and 4.6 ± 1.6 mm for RU8-15, RU8-20 and RU15-15, respectively). This increase resulted in larger follicles ( P = 0.0002) at the onset of the follicular phase (i.e. time of weaning for control sows and 24 h after last administration of Altrenogest for treated sows); 4.8 ± 1.8, 4.8 ± 1.4, 4.9 ± 0.9 mm and 2.9 ± 0.8, for RU8-15, RU8-20, RU15-15 and controls, respectively. Pre-ovulatory follicle size tended ( P = 0.07) to be larger for treated animals (7.9 ± 2.4, 7.9 ± 0.7, 8.6 ± 1.3 and 6.9 ± 0.9 mm for RU8-15, RU8-20, RU15-15 and controls, respectively). The interval follicular phase-oestrus was shorter ( P = 0.005) for treated animals. Treatment did not affect ovulation rate or early embryonic development. However, for treated animals, the increase in follicle size during treatment was positively related with ovulation rate ( P = 0.05). In conclusion, post-weaning treatment with Altrenogest of first litter sows influenced follicle size and shortened the follicular phase, but did not affect ovulation rate or early embryonic development.

439 OVARY ULTRASOUND EVALUATION OF Santa Inês EWES SUBMITTED TO SHORT DURATION PROTOCOLS WITH PROGESTOGENS ASSOCIATED WITH LECIRELIN AND/OR ESTRADIOL BENZOATE

In order to enhance the use of biotechnology in females it is necessary to have good knowledge of the ovarian events, and thus enable the manipulation of the estrus cycle to concentrate ovulations in an established period of time. In this experiment, we evaluated the ovary response and corpus luteum (CL) parameters in a short-duration protocol associated or not to lecirelin (L)(GnRH) and/or estradiol benzoate (EB). Twenty-four cyclic Santa InÊs ewes were divided into 4 groups: Control animals (G1; n = 6) received 45 μg of d-cloprostenol (Prolise®, Tecnopec, Sao Paulo, Brazil) on Day 0. Intravaginal sponges containing medroxyprogesterone acetate (MAP60®, Tecnopec) were inserted on Day 3. Then on Day 7 sponges were removed and 400 IU of eCG and 45 μg of d-cloprostenol were administered. G2 (n = 6) animals underwent the same protocol as G1 but with administration of 1 mg of EB on Day 1; G3 (n = 6) animals underwent the same protocol as G1 but with administration of 25 μg of L (Gestran plus®, Tecnopec) 30 h after sponge withdrawal; G4 (n = 6) animals underwent the same protocol as G1 but with administration of 1 mg of EB on Day 1 and 25 μg of L 30 h after sponge withdrawal. Ewe ovaries were assessed via transrectal ultrasound (US), and CL were measured 4 days after ovulation. On the same day blood samples were taken to measure plasma progesterone levels via radioimmunoassay. Twenty-three out of 24 animals ovulated. One ewe from G4 that did not ovulate was taken out of the experiment. The average size of the largest pre-ovulatory follicle was 7.3 mm (G1), 7.0 mm (G2), 6.8 mm (G3), and 7.4 mm (G4) (P &gt; 0.05). There were 1.8, 1.2, 2.0, and 1.4 ovulated follicles per animal in G1, G2, G3, and G4, respectively. The results for estrus start and duration after sponge withdrawal were G1: 32/38.6; G2: 37/69.3; G3: 29.3/37.3; G4: 44/68 h, respectively. Ovulation time span was G1: 65.3; G2: 88; G3: 53.3; G4: 82.4 h after sponge withdrawal. There was a high correlation of ovary mensuration and progesterone level (r = 0.64; P &lt; 0.0001) on the fourth day after ovulation. Progesterone levels after ovulation were 2.37 ng mL-1 (G1); 1.5 ng mL-1 (G2); 3.22 ng mL-1 (G3) and 1.99 ng mL-1 (G4), being higher in G3 than in G2 (P &lt; 0.05). It was seen that the EB is prejudicial to the CL function. Despite the fact that there was no significant difference of progesterone levels within G1, G3, and G4, animals in G3 displayed higher levels of progesterone; hence, there is a need for further studies using a larger number of animals and fertility test. CNPq e Tecnopec.

The relationship between periovulatory endocrine and follicular activity on corpus luteum size, function, and subsequent embryo survival

The objectives of this study were to examine the relationships between periovulatory endocrine events, ovarian activity, and embryo survival after artificial insemination (AI) in cattle (Bos taurus). Eighty-four reproductively normal beef heifers were estrus synchronized using a prostaglandin-based regimen. Artificial insemination was performed between 5 and 21 h after heat onset. Ultrasonic examination of ovarian structures began 12 h after the onset of heat and continued every 6 h until confirmed ovulation. Blood samples were collected for measurement of estradiol, progesterone, and insulin-like growth factor-1 (IGF-1). Pregnancy diagnosis was conducted on Days 30 and 100 after AI. Embryo survival was defined as the presence of an embryo with a detectable heartbeat in a clear amniotic sac at Day 30 postinsemination. There was no effect of the intervals from the onset of heat to AI or ovulation or from AI to ovulation on embryo survival (P > 0.10). There was a tendency (P = 0.09) of an inverse relationship between preovulatory follicle size and embryo survival that was unrelated to concentrations of estradiol or IGF-1 during the periovulatory period (P > 0.05). There was evidence (P = 0.08) of a positive association between embryo survival and concentrations of progesterone on Day 7; however, this relationship was independent (P < 0.05) of hormonal and follicular measurements during the periovulatory period. This study shows that heifers could be inseminated up to 31.5 h before ovulation without compromising the probability of embryo survival. This study suggests that there is an optimum range of follicle size within which high embryo survival rates can be achieved.

Determinants of Embryo Development and Quality in Beef Cattle: Effect of Pre-Ovulatory Follicle Size, CL Volume, and Serum Concentrations of Progesterone.

Determinants of Embryo Development and Quality in Beef Cattle: Effect of Pre-Ovulatory Follicle Size, CL Volume, and Serum Concentrations of Progesterone.

Formation of Corpora Lutea and Central Luteal Cavities and Their Relationship with Plasma Progesterone Levels and Other Metabolic Parameters in Dairy Cattle

The corpus luteum (CL) may be looked upon as a compact or cavitary structure. A number of papers have addressed the relationship between CL type and parameters such as fertility or progesterone levels. The present study assessed the morphological and functional sequence observed in cows with different CL types, comparing pre-ovulatory follicle size, progesterone levels, luteal tissue formation and some blood biochemical parameters (calcium, albumin, inorganic phosphorus, glucose, magnesium, copper and zinc), oestrus cycle length and oestrus expression, as a function of CL type. Twenty-eight lactating dairy cows from two commercial dairy farms in southern Spain were studied. Oestrus detection was performed by monitoring daily oestrus behaviour, and artificial insemination (AI) was performed using the AM/PM rule. Ovaries and uterus were sonographically examined and blood samples were collected to measure progesterone and various biochemical parameters. There was a slight tendency towards the appearance of luteal cavities when pre-ovulatory follicles were larger (1.9 +/- 0.2 vs 1.7 +/- 03; p = 0.074). Fertility was not affected by cavity presence (cavity = 42.9% and compact = 57.1%; n.s.). Luteal tissue and function were not modified as a function of CL type. Cows with luteal cavities displayed significantly higher levels of albumin, suggesting a possible metabolic influence on the formation of these structures, although specific research is required to confirm this observation.

The effect of dietary supplementation with calcium salts of long chain fatty acids and/or l-carnitine on ovarian activity of Rahmani ewes

This study investigated the effect of dietary supplementation with calcium salts of long chain fatty acids with or without of l-carnitine on ovarian activity using 24 Rahmani ewes randomly allocated to four treatments. Control animals ( n = 6) were fed a basal diet of hay (64.2%) and barley grain (35.0%) plus minerals and vitamins (0.8%). Ewes on the three treatments received the same basal diet supplemented with calcium salts of long chain fatty acids (CSFA) at 3% of the basal diet dry matter intake (1.4 kg/ewe/d); 250 ppm l-carnitine (LC); or both these supplements (CSFA + LC). All use exhibited natural estrus on one or two occasions and were weighed at the start and the end of the study as well as body condition score was assessed at the end of study. All ewes were then synchronised for estrus using intravaginal sponges for 12 d prior to the start of the nutritional treatments and three weeks after the nutritional treatments began. The nutritional treatments were imposed for a total of 8 weeks. Blood samples were collected prior to the start of treatments and every two weeks thereafter except after sponge removal of first and second synchronisation where the blood samples were collected daily for progesterone assay. The results revealed that Rahmani ewes received basal diet (control) and l-carnitine had significantly decrease final body weight and body condition score (36.3 ± 0.4; 36.8 ± 0.3; 2.2 ± 0.04; 2.1 ± 0.05; p < 0.05, respectively) than those on CSFA and CSFA + LC (38.6 ± 0.9; 39.5 ± 0.6; 3.3 ± 0.07; 3.4 ± 0.06; respectively). At the second ultrasound examination, the control animals had significantly fewer total follicles (7.3 ± 0.8; p < 0.05) than those on the CSFA (8.4 ± 0.8), l-carnitine (8.7 ± 1.5) and CSFA + LC (8.0 ± 0.6) treatments. The increased numbers occurred in the medium and large categories of follicles. In addition, the ovulation rates were significantly lower ( p < 0.05) for control (1.3 ± 0.2) and l-carnitine (1.5 ± 0.00) than for CSFA (2.5 ± 0.3) and CSFA + LC (2.3 ± 0.2). Furthermore, serum progesterone concentrations had risen and were significantly higher ( p < 0.05) for CSFA (2.5 ± 0.3 ng/ml) and CSFA + LC (2.7 ± 0.1 ng/ml) than for control (1.1 ± 0.7 ng/ml) and l-carnitine (1.5 ± 0.4 ng/ml). It was concluded that supplementation of the basal diet with l-carnitine alone did not improve performance of ewes or the ovarian response. However, the addition of calcium salts of long chain fatty acids to the basal diet alone or in combination with l-carnitine significantly improved the number and size of ovarian preovulatory follicles, and the ovulation rate of Rahmani ewes. Further evidence was required to study their influence on follicular atresia.

Pregnancy rates to timed artificial insemination in dairy cows treated with gonadotropin-releasing hormone or porcine luteinizing hormone

We compared the effects of porcine luteinizing hormone (pLH) versus gonadotropin-releasing hormone (GnRH) on ovulatory response and pregnancy rate after timed artificial insemination (TAI) in 605 lactating dairy cows. Cows (mean ± SEM: 2.4 ± 0.08 lactations, 109.0 ± 2.5 d in milk, and 2.8 ± 0.02 body condition score) at three locations were assigned to receive, in a 2 × 2 factorial design, either 100 μg GnRH or 25 mg pLH im on Day 0, 500 μg cloprostenol (PGF) on Day 7, and GnRH or pLH on Day 9, with TAI 14 to 18 h later. Ultrasonographic examinations were performed in a subset of cows on Days 0, 7, 10, and 11 to determine ovulations, presence of corpus luteum, and follicle diameter and in all cows 32 d after TAI for pregnancy determination. In 35 cows, plasma progesterone concentrations were determined 0, 3, 4, 5, 6, 7, and 12 d after ovulation. The proportion of noncyclic cows and cows with ovarian cysts on Day 0 were 12% and 6%, respectively. Ovulatory response to first treatment was 62% versus 44% for pLH and GnRH and 78% versus 50% for noncyclic and cyclic cows ( P < 0.01). Location, ovulatory response to first pLH or GnRH, cyclic status, presence of an ovarian cyst, and preovulatory follicle size did not affect pregnancy rate. Plasma progesterone concentrations after TAI did not differ among treatments. Pregnancy rate to TAI was greater ( P < 0.01) in the GnRH/PGF/pLH group (42%) than in the other three groups (28%, 30%, and 26% for GnRH/PGF/GnRH, pLH/PGF/GnRH, and pLH/PGF/pLH, respectively). Although only 3% of cows given pLH in lieu of GnRH on Day 9 lost their embryo versus 7% in those subjected to a conventional TAI using two GnRH treatments, the difference was not statistically significant. In summary, pLH treatment on Day 0 increased ovulatory response but not pregnancy rate. Cows treated with GnRH/PGF/pLH had the highest pregnancy rate to TAI, but progesterone concentrations after TAI were not increased. In addition, preovulatory follicle diameter did not affect pregnancy rate.

Effects of additional prostaglandin F2α and estradiol-17β during Ovsynch in lactating dairy cows

This study was designed to evaluate whether decreasing circulating progesterone (P4) or increasing circulating estradiol-17β (E2) near the time of artificial insemination (AI) in an Ovsynch protocol would increase pregnancies per AI (P/AI) in lactating dairy cows. Six hundred nineteen lactating Holstein cows (n=772 inseminations) received Ovsynch (GnRH–7 d-PGF2α–56 h-GnRH–16 h-timed AI). Cows were randomized in a 2×2 factorial experiment of 4 treatments to receive or not receive 25mg of PGF2α 24h after the standard PGF2α of Ovsynch, or 0.5mg of E2 at the time of the final GnRH of Ovsynch, or both. Blood samples were collected 24h after normal PGF2α and at final GnRH to evaluate circulating P4. Ovarian ultrasound was done at final GnRH to determine preovulatory follicle size. Ovulation was confirmed by ultrasound 5 d after AI. Treatment with additional PGF2α increased the percentage of cows that had complete luteal regression (95.6%) compared with control cows (84.6%). In contrast, additional PGF2α had no detectable effect on P/AI (control = 41.5% vs. + PGF2α=44.7%). Supplementation with E2 increased expression of estrus (84.4 vs. 37.2%), but had no effect on overall fertility and even tended to have a negative effect on fertility in cows that ovulated to the second GnRH (control = 51.5% vs. +E2=44.0%). Thus, additional treatments with PGF2α or E2 during Ovsynch can be used to increase synchronization and expression of estrus during Ovsynch, although the lack of improvement in fertility makes these treatments unwarranted.

The study of the ovaries' state in local Bulgarian ewe's breeds by real-time ultrasonography

The aim of this study was the characterization of the ovaries' state in local Bulgarian ovine breeds at the moment of ovulation by real-time transrectal ultrasonography. It was observed 32 ewes of Bulgarian local breeds Karakachanska and West-Oldmountainous between 1,5 and 4 years old, housed under closely to their natural conditions. Ovaries were examined ultrasonically after estrus synchronization with the implantation of intravaginal sponges containing 45 mg of Cronolone over 14 days. The effect of age and breed on the variability of the ovary's size and preovulatory follicle's size was studied.

Preovulatory Follicle Development in Goats Following Oestrous Synchronization with Progestagens or Prostaglandins

The study reports on differences in the dynamics of growth and functionality of preovulatory follicles in response to oestrous synchronization, either by the administration of two doses of prostaglandin or by an intravaginal progestagen sponge, in goats. The progestagen-treated group (n = 8) showed more follicles of preovulatory size (> or =5.5 mm) than the cloprostenol group (n = 8) during the follicular phase (4.5 +/- 0.6 vs 1.9 +/- 0.2, p < 0.01). The diameters of the largest follicles (LF1, LF2 and LF3) were also larger in the progestagen group (LF1, 7.8 +/- 0.3 vs 7.0 +/- 0.2 mm, p < 0.05; LF2, 6.7 +/- 0.2 vs 5.6 +/- 0.2 mm, p < 0.01; LF3, 5.5 +/- 0.3 vs 4.2 +/- 0.2 mm, p < 0.01). The study of the preovulatory follicles showed that 27.2% (3/11) of the follicles were in the static phase in the cloprostenol group, whilst 71.4% (10/14) were static in progestagen group (p < 0.05). Higher plasma oestradiol levels were recorded in the progestagen-treated goats during the 48 h prior to cloprostenol injection or progestagen withdrawal (4.2 +/- 0.4 vs 3.0 +/- 0.2 pg/ml, p < 0.05). In conclusion, goats with oestrus synchronized by progestagen showed a higher number of preovulatory-sized follicles, but a decreased oestradiol secretion when compared with does with oestrus synchronized by using prostaglandin analogues. These would support the development of alternative protocols for assisted reproduction.

Superovulatory Ovarian Response in Mangalica Gilts is Not Influenced by Feeding Level

The aim of the study was to compare how different feeding levels affect the ovarian potential of follicular development and oocyte maturation in response to superovulatory treatment in native Mangalica (M, n = 17) compared with Landrace (L, n = 20) pigs. Gilts of both breeds were fed high-energy (HI-2.5 kg) or low-energy (LO - 1.25 kg) feed during oestrus synchronization (15 days of Regumate feeding) till the time of oocyte aspiration (Day 6 after Regumate). Follicular growth was stimulated by the administration of 1000 IU equiue choriou gonadotropiu (eCG) 24 h after Regumate treatment, and ovulation was induced by injection of 750 IU human choriou gonadotropiu (hCG) 80 h after eCG administration. Ultrasound (US) investigation was done three times (4-10 h before, and 40-44 and 72-74 h after eCG administration) for the observation of follicular development. Oocyte and follicular fluid (FF) were collected endoscopically 34 h after hCG injection. Cumulus-oocyte complexes were evaluated, their morphology determined, and thereafter fixed and stained for chromatin evaluation. Oocytes were classified as meiosis-resumed (germinal vesicle breakdown, diakinesis, metaphase I to anaphase I) or matured (telophase I and metaphase II). FF concentrations of oestradiol and progesterone were measured by validated radioimmunoassays. In L gilts, differences were observed between HI and LO in the number of preovulatory follicles (32.3 +/- 10.5 vs 17.1 +/- 12.3, p < 0.05), but not in M (25.3 +/- 2.9 vs 28.8 +/- 7.3, p > 0.05). Initial follicular growth was not affected by feeding levels; however, preovulatory follicle size was larger in M (7.1 +/- 0.9 and 6.9 +/- 1.1 mm vs 5.7 +/- 0.7 and 5.5 +/- 0.8 mm; p < 0.05). No differences were obtained with relation to mature chromatin configuration in both breeds (L gilts: HI - 70% and LO-67% vs M gilts: HI - 67% and LO - 63%). A twofold higher oestradiol concentration was detected in FF of HI-M and LO-M (29.6 +/- 6.8 and 30.9 +/- 10.3 ng/ml respectively) compared with that of L (16.9 +/- 9.7 and 17.9 +/- 3.6 ng/ml, respectively; p < 0.05). The mean FF progesterone level was nearly fivefold higher in M (2020.4 +/- 1056 and 1512.2 +/- 1121.8 ng/ml) compared with L (386.2 +/- 113.7 and 298.8 +/- 125.9 ng/ml, p < 0.05). The results indicate an influence of the feeding of altered energy on the number of recruitable preovulatory follicles in modern Landrace but not in native Mangalica breed. Moreover, the follicular steroid hormone milieu differs between Landrace and Mangalica gilts but not depending on feeding levels. Oocyte maturation was not affected by diet.