Number Of Medium-sized Follicles Research Articles

Radioactive Iodine-induced hypothyroidism interferes with the maturation of reproductive organs during puberty in immature female rats.

Animal and human studies suggest that thyroid hormone may have critical roles in the development of the ovary. For example, thyroid deficiency disrupts the ovarian microarchitecture and menstrual cycle in neonate and adult women, respectively. Therefore, it is conceivable that thyroid deficiency might disrupt sexual maturation during the peri-pubertal period. To investigate the impact of radioactive iodine-induced thyroid deficiency on reproductive organs throughout puberty, immature female rats were given water containing radioactive iodine (0.37MBq/g body weight) twice, on postnatal days 22 and 29. Radioactive iodine-induced hypothyroidism was revealed by low free thyroxin levels. Thyroid deficiency delayed the onset of vaginal opening, reduced ovarian weight and the number of medium-sized follicles and led to elongated uteri. However, there was no effect on the estrous cycle or absolute uterus weight. We conclude that radioactive iodine-induced thyroid deficiency delays sexual maturation and alters normal ovarian growth in peri-pubertal rats.

Effect of three schemes of ovum pick-up on the follicular dynamics, gene expression, and in-vitro developmental competence of oocytes in Sahiwal cattle.

This study aimed to compare the effect of three schemes of ovum pick-up (OPU) on follicular dynamics, oocytes recovery, oocytes quality, gene expression, nuclear maturation and in-vitro developmental competence of oocytes in Sahiwal cattle. Considering the follicle population, all the cows were divided equally in a 3× 3 cross over design, and each cow received one of the three treatments: (a) twice weekly (TW; n=6), (b) once weekly (OW; n=6) and (c) bi-weekly OPU (BW; n=6) in three periods, with the first OPU conducted on 4, 7 and 14 days after second dominant follicle puncture (DFP) in the TW, OW and BW OPU interval groups, respectively. The collected cumulus oocytes complexes (COCs) were graded into A, B, C and D grades depending on the number of layers of cumulus cells and homogeneous nature of cytoplasm. Nuclear maturation was assessed by staining the oocytes with Hoechst 33342. The growth rate (mm/day) of dominant follicle (DF) (F1) (0.49 ± 0.21 vs. 0.71 ± 0.26 vs. 1.30 ± 0.27) and first subordinate follicle (F2) (0.85 ± 0.27 vs. 0.71 ± 0.25 vs. 1.06 ± 0.29) did not differ (p > .05) among all the three groups. The proportion of animals bearing a corpus luteum (CL) in the BW OPU interval group (53.3%) was significantly higher (p < .05) as compared to TW (13.3%) and OW (18.3%) OPU interval groups. The number of medium-sized follicles and oocyte with grade A and B were significantly higher (p < .05) in the TW (1.16 ± 0.21 and 33.88 ± 0.03) OPU interval group as compared to the OW (0.88 ± 0.22 and 21.54 ± 0.03) and BW (0.55 ± 0.21 and 21.89 ± 0.02) OPU interval groups. However, the number of degenerated oocytes in BW (0.85 ± 0.16) OPU interval group was significantly higher (p < .05) as compared to the TW (0.16 ± 0.15) and OW (0.44 ± 0.16) OPU interval groups. Expression level of growth differentiation factor 9 in TW OPU interval group was significantly higher (p < .05) as compared to the OW and BW OPU interval groups. Likewise, expression level of bone morphogenetic protein 15 (BMP15) in the TW and BW OPU interval groups was significantly higher (p < .05) as compared to the OW OPU interval group. The nuclear maturation rate was significantly higher in the TW (63.64 ± 0.07) and BW (59.26 ± 0.08) OPU groups as compared to OW (51.43 ± 0.06) OPU interval group. However, the cleavage rate (59.30 ± 0.06 vs. 44.29 ± 0.06 vs. 56.67 ± 0.06) did not differ (p > .05) among the three groups. Whereas, the blastocyst rate tended to be higher (p=.06) in the TW (29.07 ± 0.05) and BW (28.33 ± 0.04) OPU interval groups as compared to OW (18.57 ± 0.05) OPU interval group. Taken together, it can be concluded that TW OPU interval scheme enhances the medium-sized follicles resulting in good quality oocytes, regulates the oocyte-derived paracrine factors, leading to higher nuclear maturation rates and improved embryonic development in-vitro.

Synchronization of follicle wave emergence before ovarian superstimulation with FSH and ovum pick-up improves in vitro embryo production in pregnant heifers

The objective of the present study was to evaluate effects of synchronization of timing of follicle wave emergence, before ovarian superstimulation and ovum pick-up (OPU), on ovarian response and embryo production in pregnant heifers. Pregnant (47–69 days of gestation) Holstein heifers (n = 64), 19.0 ± 0.3 months of age, were assigned in a completely randomized design to one of two groups: synchronization of follicular wave emergence using follicle ablation (Synchronized) or untreated control (Non-synchronized). Superstimulatory treatments consisting of 160 mg (280 IU) of porcine follicle stimulating hormone (p-FSH), administered in four decreasing dose treatments 12 h apart, were initiated 36 h after follicle ablation or at random stages of the follicular wave in heifers of the Synchronized and Non-synchronized group, respectively. Ovum pick-up was performed in all heifers 40 h after the last p-FSH administration and retrieved cumulus oocyte complexes (COCs) were subjected to in vitro embryo production (IVEP) procedures. Ultrasonography was performed immediately before OPU to determine number and size of ovarian follicles. Differences in treatment responses between groups were evaluated using generalized linear mixed models. Total number of follicles at the time of OPU was not different between treatment groups (P = 0.61), however, the number of small follicles (<6 mm) was greater (P = 0.05) in heifers of the Non-synchronized group, whereas number of medium size follicles (6–10 mm) tended (P = 0.09) to be greater in heifers of the Synchronized group. Heifers in the Synchronized group had a greater (P = 0.01) percentage of medium sized follicles and a lesser (P = 0.01) percentage of smaller sized follicles than heifers in the Non-synchronized group. There were no differences (P > 0.15) in total number of recovered COCs, or number of viable COCs between groups. Cleavage percentage (84.5% and 72.8%) and blastocyst percentage (48.2% and 33.4%) were greater (P < 0.01) in heifers of the Synchronized than Non-synchronized group, respectively. As a result, mean number of blastocysts per OPU/heifer was greater (P = 0.006) in the heifers of the Synchronized (8.9 ± 1.0) than the Non-synchronized (5.5 ± 0.9) group. In conclusion, synchronizing the time of follicle wave emergence in pregnant heifers, prior to ovarian superstimulation with FSH and OPU results in a greater superstimulatory response and oocyte competence leading to greater embryo production.

71 Effect of follicle-stimulating hormone dose and circulating progesterone before ovum pickup and invitro embryo production in pregnant Holstein heifers

Superstimulation with exogenous FSH before ovum pickup for invitro embryo production (IVP) improves embryo production. However, there is limited information on different FSH dose regimens and the effect of progesterone (P4) concentrations on IVP. Thus, the objective of the present study was to evaluate the effect of FSH dose and circulating P4 on ovarian superstimulation responses, before ovum pickup (OPU) on IVP in pregnant heifers. Pregnant Holstein heifers (n=47) 19.5±0.3 months of age with moderate body condition score (3.3±0.03; scale 1 to 5) were randomly assigned to 2 treatment groups, p-FSH (Folltropin-V, Vetoquinol) 160mg (FSH160) or p-FSH 300mg (FSH300), in a crossover design. Two sessions of OPU were performed on each animal at 48 and 62 days of gestation, with a washout interval between sessions of 14 days. Dominant follicle removal (DFR) was utilised to synchronize follicular wave emergence, and FSH treatments were initiated 36h later. Heifers in both groups received a total of 4 FSH injections 12h apart as follows: FSH160 (48.0, 42.7, 37.3, and 32.0mg) or FSH300 (90.0, 80.0, 70.0, and 60.0mg). All heifers underwent OPU 40h after the last FSH treatment. Blood samples were collected for P4 quantitation at OPU and number of small (&amp;lt;6mm), medium (6–10mm), and large (&amp;gt;10mm) follicles were determined before OPU. Cumulus–oocyte complexes (COCs) recovered were processed and invitro embryo production was performed using standard procedures. Differences between treatment groups were evaluated using generalized linear mixed models, and data are presented as mean±s.e.m. (Table 1). Heifers treated with 300mg of p-FSH had a greater (P&amp;lt;0.05) number of medium-size follicles, total number of follicles, total COCs recovered, number of viable COCs, and number of blastocysts produced. In addition, heifers treated with 300mg of p-FSH had a greater cleavage rate (P=0.03). Circulating P4 was on average 10.1±0.4ng mL−1 and did not differ between gestation days (P=0.12). Furthermore, circulating P4 did not affect (P&amp;gt;0.10) any of the outcomes evaluated, nor was there an FSH dose by circulating P4 interaction (P&amp;gt;0.05). In conclusion, increasing the dose of p-FSH before OPU from 160 to 300mg resulted in an increase in the total number of follicles, number of oocytes, and number of blastocysts produced in pregnant donors without changes in blastocyst rate, regardless of progesterone concentrations at the time of OPU. Table 1. Ovum pickup and invitro embryo production in pregnant heifers treated with different doses of p-FSH Item FSH160 FSH300 P-value Small follicles (n) 5.0±0.6 3.5±0.4 0.02 Medium follicles (n) 14.4±0.8 18.3±0.9 0.0003 Large follicles (n) 2.9±0.3 2.5±0.3 0.28 Total follicles (n) 22.3±0.7 24.3±0.9 0.04 Total oocytes (n) 12.8±0.7 14.7±1.0 0.02 Recovery rate (%) 56.2±2.1 59.7±2.7 0.20 Viable oocytes (n) 11.4±0.7 13.3±1.0 0.02 Viable oocytes (%) 89.9±1.7 88.0±2.4 0.22 Cleavage rate (%) 63.8±4.3 71.5±3.7 0.03 Cleaved oocytes (n) 7.2±0.7 9.3±0.9 0.14 Blastocysts (n) 2.7±0.4 3.6±0.7 0.02 Blastocyst rate (%) 22.0±3.3 22.4±3.6 0.74

120 Effects of antral follicle count in ovaries on follicular development and endocrine dynamics of follicle-stimulating hormone and steroid hormones in cattle

The antral follicle count (AFC) in mammalian ovaries positively correlates with female fertility. We have reported previously that cumulus-oocyte complexes derived from high-AFC ovaries produce more oestradiol-17β (E2) and have higher developmental competence than those from low-AFC ovaries in in vitro growth (Sakaguchi et al. WCRB2017, Okinawa, Japan; 1-15). We also indicated that follicular growth was different between high- and low-AFC cows, and that follicular deviation occurred earlier in high-AFC cows (Nagai et al. 2015 Anim. Reprod. Sci. 163, 172-178). To clarify the causes of the higher E2 production and earlier follicular deviation in high-AFC cows, we investigated circulating steroid hormones and FSH during the oestrous cycle, as well as steroidogenesis in follicles at different phases of the oestrous cycle. Four high-AFC cows (&amp;gt;40 follicles) and 3 low-AFC cows (&amp;lt;20 follicles) based on the peak AFC were used. We ablated their follicles 8 days after oestrus and administered prostaglandin F2α on Day 12. After 40h of prostaglandin F2α injection, follicular fluid of a dominant follicle (preovulatory phase) was collected and subordinate follicles were ablated. Gonadotropin-releasing hormone was then injected to induce LH surge. Seven days after gonadotropin-releasing hormone, follicular fluids were collected from dominant follicles (luteal phase) and all follicles were ablated. Four days later, follicular fluids were again collected from the largest follicles (selection phase). Steroid hormones in follicular fluids were measured by enzyme immunoassay. Throughout the study, we daily examined ovaries by ultrasonography and collected blood for hormone measurements. We analysed data using two-way ANOVA followed by Tukey-Kramer or Student’s t test. The number of small follicles (&amp;lt;4mm) was greater in high-AFC than low-AFC groups (low v. high=7.6v. 46.9; P&amp;lt;0.05), whereas that of large follicles (&amp;gt;8mm) was similar between groups (0.8v. 0.8) at any time. Although the number of medium-sized follicles (4-8mm) was not affected in the low-AFC group (4.2-6.7) after follicle ablation, the number increased from 1 to 4 days (10.5 to 17.8) then decreased 6 days after follicle ablation (11.5) in the high-AFC group, which means that degradation of follicles occurs at selection phase in high-AFC cows. Peripheral FSH concentration was higher (low v. high: 4.7v. 2.1 ng/mL), but E2 concentration was lower in the low-AFC (2.4 pg/mL) than high-AFC group (3.3 pg/mL) during the selection phase (P&amp;lt;0.05). Although follicular diameter was similar in both groups at any phase, E2 (low v. high=333.8v. 1127.4 ng/mL; P&amp;lt;0.05) and testosterone (low v. high=16.7v. 35.0 ng/mL; P=0.07), but not progesterone, concentrations were higher in the high-AFC than low-AFC group in the preovulatory phase. These findings suggest that a lower response to FSH causes low E2 production in the low-AFC group, resulting in high FSH concentration and a consistent development of medium-sized follicles. Conversely, higher E2 concentration suppresses FSH secretion, resulting in obvious degradation of follicles in the high-AFC group at the selection phase.

Experimental model of toxin-induced subclinical mastitis and its effect on disruption of follicular function in cows

This study establishes an experimental model for subclinical mastitis induced by Gram-positive (G+) exosecretions of Staphylococcus aureus origin or Gram-negative (G−) endotoxin of Escherichia coli origin to examine its effects on follicular growth and steroid concentrations in Holstein dairy cows. Cows were synchronized with the Ovsynch protocol followed by a series of follicular cycles that included GnRH and PGF2α doses administered every 8 days. Cows received small intramammary doses of either G+ (10 μg, n = 10) or G− (0.5 μg, n = 6) toxin, or saline (n = 6; uninfected control) every 48 hours for 20 days. Follicular fluids were aspirated from preovulatory follicles before (aspiration one: control), at the end of (aspiration two: immediate effect), and 16 days after the end of (aspiration three: carryover effect) toxin exposure. During the 3 weeks of subclinical mastitis induced by G+ or G−, no local inflammatory signs were detected in the mammary gland and no systemic symptoms were noted: body temperatures of the treated cows did not differ from controls; plasma cortisol and haptoglobin concentrations were not elevated and did not differ among groups. Somatic cell count was higher in the treated groups than in controls, and higher in the G− versus G+ group. For analysis of reproductive responses, cows were further classified as nonaffected or affected based on an more than 20% decline in follicular androstenedione concentration in aspiration two or three relative to the first, control aspiration. Most G− (5/6) and 40% of G+ (4/10) cows were defined as affected by induced mastitis. An immediate decrease in the number of medium-size follicles was recorded on Day 4 of the induced cycle, toward the end of the 20-day mastitis induction, in the affected G+ compared with uninfected control group (1.0 ± 0.5 vs. 3.0 ± 0.4 follicles; P < 0.05); the affected G− and nonaffected G+ subgroups exhibited a similar numerical decline in the number of follicles. A carryover (but not immediate) decrease to 51% and 62% in follicular estradiol concentrations in G− affected group and G+ affected group was detected relative to controls (P < 0.05). The nonaffected G+ subgroup did not differ from its control counterparts. Based on the current experimental model, subclinical IMI induced by G+ or G− toxin disrupts follicular functions, and it seems that the ovarian pool of early antral follicles is susceptible to subclinical mastitis.

Fever in women may interfere with follicular development during controlled ovarian stimulation

Purpose: This study aims to evaluate the effects of fever on follicular development in women undergoing controlled ovarian stimulation during in vitro fertilisation (IVF) and intracytoplasmic sperm injection (ICSI) cycles.Materials and methods: This was a retrospective observational self-controlled study at a tertiary-care fertility centre. Six gonadotropin stimulation cycles characterised by poor ovarian response in which women reported the occurrence of a febrile illness, were considered for evaluation. Fever-exposed cycles were compared to the next stimulation cycle in the same women. Primary outcome measures were final number of pre-ovulatory follicles (≥ 16 mm) and final peak serum estradiol levels (pg/mL). Other outcome measures were final number of medium-sized follicles (12–15 mm), final mean estradiol serum level per follicle ≥ 12 mm (pg/mL), total days of stimulation and total gonadotropin ampoules utilised.Results: Fever-exposed cycles were associated with significantly lower number of pre-ovulatory follicles (0.7 ± 0.8), significantly higher number of medium-size follicles (21.0 ± 4.5), and significantly reduced serum estradiol per follicle ≥12 mm (50.7 ± 11.7 pg/mL). They also required a significantly longer duration of ovarian stimulation (15.7 ± 3.3 days) and a significantly increased number of gonadotropin ampoules (47.2 ± 10.9). Four women had polycystic ovary syndrome and one hypothalamic hypogonadism.Conclusion: This preliminary report suggests a possible negative effect of fever on follicular development and ovarian estradiol production in some women undergoing controlled ovarian stimulation.

Maturation Competence of Swamp Buffalo Oocytes Obtained by Ovum Pick‐Up and from Slaughterhouse Ovaries

This study was designed with the final goal of improving in vitro embryo production in the Thai swamp buffalo (Bubalus bubalis carabensis). Oocytes were collected by ovum pick-up (OPU) from six non-lactating multiparous swamp buffalo twice per week for 10 consecutive sessions followed by once-weekly collection for 10 consecutive sessions without hormone stimulation. In addition, oocytes were collected from slaughterhouse ovaries that were classified as follows: ovaries from non-pregnant cows with a visible corpus luteum (NPCL); pregnant cows with a corpus luteum (P); and non-pregnant cows without a corpus luteum (NP). Follicles in each group of ovaries were categorized as small (2-4 mm), medium-sized (5-8 mm) or large follicles (≥ 9 mm). The quality of the oocytes was assessed by their capacity to undergo in vitro maturation. The total number of observed follicles per session (all sizes combined) was larger in the once-weekly OPU group compared with the twice-weekly OPU group. In particular, the numbers of small and large follicles were higher in the once-weekly OPU group (5.2 ± 0.7 and 0.9 ± 0.2, respectively) than in the twice-weekly OPU group (3.9 ± 0.5 and 0.5 ± 0.1). The number of medium-sized follicles did not differ between the groups. The percentages of oocytes with an abnormal spindle morphology were not different between oocytes from the twice-weekly (30.0%) and the once-weekly (28.6%) OPU groups. A higher percentage of oocytes obtained in vitro (49.5%) exhibited nuclear abnormalities compared with those obtained in vivo (≤34.8%) after in vitro maturation. In conclusion, oocytes can be successfully collected by OPU in the swamp buffalo, without hormonal pretreatment, and per week more good-quality oocytes can be collected by twice-weekly OPU. In addition, oocytes collected from slaughterhouse ovaries can be used with the reproductive status of the cow having no influence on the maturation competence of oocytes.

Follicular characteristics and intrafollicular concentrations of nitric oxide and ascorbic acid during ovarian acyclicity in water buffalo (Bubalus bubalis)

The objective of this study was to examine the follicular characteristics and intrafollicular concentrations of nitric oxide and ascorbic acid during ovarian acyclicity in buffaloes. Ovaries were collected from 56 acyclic and 95 cyclic buffaloes at slaughter, surface follicle number was counted and follicles were classified into small (5.0-6.9 mm), medium (7.0-9.9 mm), and large (≥ 10.0 mm) size categories based on their diameter. Follicular fluid was aspirated and assayed for nitric oxide, ascorbic acid, estradiol, and progesterone. Acyclic buffaloes had a higher (P<0.05) number of medium-sized follicles and a lower (P<0.001) number of large follicles than the cyclic ones. In acyclic animals, the number of large follicles was lower (P<0.01) than in medium size category which in turn was lower (P<0.001) than the number of small follicles. In contrast, the number of medium and large follicles was not different (P>0.05) in the cyclic control. However, the number of small-sized follicles was higher (P<0.001) compared to the other two categories. The incidence of large-sized follicles was lower (P<0.05) in acyclic buffalo population compared to the cyclic control. Evaluation of estrogenic status demonstrated that all the follicles of acyclic buffaloes are estrogen-inactive (E (2)/P (4) ratio<1). Small- and medium-sized follicles of acyclic buffaloes had higher concentrations of nitric oxide (P<0.05 and P<0.001, respectively) and lower concentrations of ascorbic acid (P<0.05 and P<0.01, respectively) than the corresponding size estrogen-active follicles of their cyclic counterparts. In conclusion, this study indicates that follicular development continues during acyclicity in buffaloes. Although follicles in some acyclic buffaloes attain a size corresponding to morphological dominance, they are unable to achieve functional dominance, perhaps due to an altered balance of intrafollicular nitric oxide and ascorbic acid and, as a result, these follicles instead of progressing to ovulation undergo atresia.

Women, weight, and fertility: The effect of body mass index on the outcome of superovulation/intrauterine insemination cycles

To determine the fecundity of overweight and obese infertile women treated with gonadotropins and undergoing intrauterine insemination (IUI). Retrospective study. Academic infertility clinic. Four hundred seventy-seven women undergoing 1,189 ovulation induction (OI)/IUI cycles stratified by body mass index (BMI). None. BMI groups were compared regarding the following: gonadotropin dose, duration of treatment, peak E(2), number of follicles (total, large, and medium size), E(2)/follicle, endometrial thickness, spontaneous abortion, and clinical and multiple pregnancy rates. There was a significant trend toward higher medication requirements and lower E(2) levels with increasing BMI. BMI was inversely associated with [1] the E(2) level per produced preovulatory follicle and [2] the number of medium-size follicles. Furthermore, BMI was inversely associated with the number of medium, large, and total follicles divided by total FSH dose, suggesting that women with a higher BMI develop a lower number of medium and/or large follicles at a given total FSH dose. BMI was positively associated with endometrial thickness, and endometrial thickness was positively associated with pregnancy. Mean number of cycles required to conceive, clinical pregnancy, and spontaneous abortion rates did not differ significantly among the different BMI categories. Obese women require higher doses of medication and produce fewer follicles for a given dose, but once medication and response are adjusted to overcome the weight effect, the success of the treatment cycle is comparable to that of normal weight women.

Reproductive Responses to Daily Injections with Porcine Somatotropin Before Mating in Gilts

Litter size and progeny birth weights are lower in gilts than in sows. Somatotropin (ST) is an important regulator of ovulation, fetal growth and survival. We therefore investigated effects of pST treatment of gilts for two to four weeks before mating on ovulation rate, behavioural estrus, fetal growth and survival, litter size and birth weights. In Experiment One, gilts were injected with 0, 30, 60 or 90 µg pST/kg/day for 14 days commencing 7 days after first estrus. Reproductive tracts were collected and corpora lutea and follicle numbers counted 5.5 days after second estrus. Ovulation rate (P=0.031) and number of medium-sized follicles (P=0.059) correlated positively with pST dose. In Experiment Two, gilts were injected with 0, 12.5, 25 or 50 µg pST/kg/day for 21 days from first estrus, and mated at second estrus. Numbers of corpora lutea, follicles and fetuses were counted at day 31 of pregnancy. Numbers of medium follicles and ovary weights were positively related to pST dose. In Experiment Three, 31 week old (1(st) replicate) or 27 week old (2(nd) replicate) gilts were injected daily with 0 or 12.5 µg pST/kg/day until mating 25.9 ± 0.6 days later, and delivered at term. Pre-mating pST increased total litter size in younger gilts in the 2(nd) replicate only (P<0.05). In conclusion, injecting gilts with pST before mating does not consistently alter ovulation rate, increases the number of medium follicles available for recruitment at the second mating after treatment and increases subsequent litter size in younger gilts.

Effect of exogenous administration of buffalo follicular fluid on follicular development, estrus response and luteal function in anoestrous goats ( Capra hircus)

The effect of buffalo follicular fluid (buFF) on follicular development, estrus response and luteal function was investigated in anoestrous does. Treatment with buFF (18 ml/doe) had no significant effect on the number of antral follicles of all class categories during the period of administration. However, after cessation of buFF treatment, the number of total antral follicles increased significantly with time ( P < 0.003) as well as due to the treatment × time interaction ( P < 0.02), without any influence on follicle size. Injection of buFF also caused a marked increase ( P < 0.049) with time in the number of medium-sized follicles at cessation. Approximately 60 and 20% of buFF-treated anoestrous does showed behavioural and silent estrus, respectively, compared to none in the control. The mean interval between cessation of buFF treatment to onset of oestrus and oestrus duration was 67.0 ± 18.5 and 17.0 ± 3.6 h, respectively. Corpus lutea size varied between 4.6 and 5.8 mm with an average diameter of 5.2 ± 0.3 mm. Only 33.3% of does showed serum progesterone levels above 1 ng/ml, while the remainder (66.7%) had below 0.5 ng/ml. Our results indicate that exogenous administration of buFF causes enhanced follicular activity following cessation of treatment, which results in behavioural oestrus and corpus luteum (CL) development in anoestrous does. CL development and its function is, however, inadequate in buFF-treated anoestrous does.

Nutrition, Metabolism, and Fertility in Dairy Cows: 2. Dietary Fatty Acids and Ovarian Function

Plasma insulin has important implications for ovarian function in dairy cows. Previous work demonstrated that plasma insulin increased with increasing dietary starch and decreasing dietary fatty acid concentrations. The objective of this experiment was to investigate hormonal and ovarian responses to dietary fatty acid content with no change in other dietary components. Thirty cows were fed a standard diet from calving until 40 d in milk (DIM) and then 6 cows were transferred to each of 5 diets containing 0, 8, 15, 23, and 30 g/kg of dry matter (DM) of calcium salts of palm fatty acids (CaPFA; Megalac) until 70 DIM. Estrus was synchronized at 60 DIM. Between 60 and 70 DIM, energy intake, milk yield, and energy balance were similar among diet groups. Plasma insulin decreased when dietary concentration of CaPFA exceeded 15 g/kg of DM (insulin: 0.46, 0.41, 0.46, 0.33, 0.28±SE 0.034 ng/mL for diets containing 0 to 30g of CaPFA/kg of DM, respectively). Maximum plasma insulin to glucagon ratio was observed with 15g of CaPFA/kg of DM (ratios: 3.99, 4.33, 4.67, 3.45, 2.89±SE 0.156 for diets containing 0 to 30g of CaPFA/kg of DM, respectively). Plasma concentrations of growth hormone, insulin-like growth factor-I and leptin did not vary between diets. The number of small (<5mm) ovarian follicles was negatively related to plasma insulin concentration (r = −0.328) and was stimulated by CaPFA supplementation at all rates tested compared with cows receiving zero CaPFA (small follicles preovulation: 6.7, 11.2, 11.5, 11.3, 11.9±SE 1.48 for diets containing 0 to 30g of CaPFA/kg of DM, respectively). The number of medium-sized follicles, and diameters of the ovulatory follicles and corpora lutea, were not affected by CaPFA supplementation. It is concluded that dietary total fat concentration should be below 50 g/kg of DM to avoid depressing plasma insulin concentration in cows at the start of the breeding period.

Is rapid follicular development during controlled ovarian hyperstimulation (COH) for IVF and ICSI detrimental to success rate?

Objective: To determine whether a rapid follicular growth pattern and a short follicular phase (SFP) during COH in IVF and ICSI cycles is associated with certain clinical or biochemical characteristics of such women, and whether it has an effect on the outcome of such cycles. Design: Retrospective controlled study. Materials/Methods: All IVF and ICSI cycles performed between 1–1-1998 and 31–12–2001 were reviewed. Cycles in which >=3 follicles had reached >=17 mm diameter after only nine days of COH and on which oocytes retrieval was performed on day 11 of cycle were identified as cycles with rapid follicular development and SFP. This was the study group of interest. Cycles in which COH was initiated on the same day as each of SFP cycles and in which oocytes retrieval was performed on cycle day 14 or 15 served as the control group. Cycles using donated oocytes were excluded. All women underwent pituitary down regulation using a short acting GnRH agonist followed by follicle stimulating hormone (FSH) or human menopausal gonadotrophin (HMG). Results: The study included 54 SFP cycles and 169 control cycles. Women age, type of infertility (primary or secondary) or order of IVF and ICSI cycles (first or repeat cycle) were not significantly different between SFP group and control group. Equally, basal FSH levels, treatment type (IVF or ICSI), total dose or type of used FSH or HMG were not significantly different. Male factor infertility was slightly more common in control group than in SFP group 92 (54%) vs. 20 (37%). There were no significant differences in other causes of infertility (tubal, anovulation, poor ovarian reserve, endometriosis and unexplained). The number of large size follicles (>=17mm) (5.4 vs. 5.1) or the total number of large and medium (14–16mm) size follicles (8.6 vs. 9.6) on ultrasound scan 48 hours prior to oocytes retrieval were not significantly different. There were less medium size follicles in SFP group (3.2 vs. 4.5, P <0.01). The number of retrieved oocytes (8.1 vs. 9.6, p = 0.03) and the number of fertilized oocytes (5.3 vs. 6.7, P = 0.04) were less in the SFP group. However, there was no significant difference in fertilization rate (70% vs. 69.7%), number of transferred embryos (2.1 vs. 2.05) and number of frozen embryos (2.1 vs. 2.8). The clinical pregnancy rate [18 (33%) vs. 63 (37%)], ectopic pregnancy, miscarriage or live birth rates were not significantly different between the two groups. Conclusions: In our study a SFP was not associated with advanced women’s age or higher basal FSH levels. The smaller number of medium size follicles in the SFP group suggests that a second cohort of follicles did not have a chance to complete its development. This reflected itself in a smaller number of retrieved and fertilised oocytes. Nevertheless, the numbers of frozen embryos, clinical pregnancy, live birth rates were not different and the clinical significance of these differences is therefore doubtful. A SFP seems therefore to have no detrimental effect on oocyte quality, endometrial development or embryo implantation. Although a SFP in the spontaneous cycle has been found by some to be associated with reduced fertility, this appeared not to be the case during COH for IVF and ICSI. During COH the outcome of cycles with SFP is comparable to cycles with ‘normal’ follicular phase length. Supported by: Centre for Reproductive Medicine, Coventry, UK.

Alteration of follicular dynamics and superovulatory responses by gonadotropin releasing hormone and follicular puncture in cattle: A field trial

A field experiment was conducted to determine the influence of follicular alteration on superovulatory responses. Ultrasonography was performed once daily over 4 d prior to gonadotropin treatment (Day 0), on the day of estrus during superstimulation, and on the day of embryo collection to monitor follicular development. Animals were superstimulated between Days 8 and 12 of the estrous cycle. Follicular status was altered 2 d prior to initiation of superstimulation (Day 0) with GnRH (Cystorelin, 200 μg im) administered with (GnRHpuncture group, n=31) or without (GnRH-no puncture group, n=52) concomitant removal of the largest follicle by follicular aspiration. Responses were compared with those of an untreated control group superovulated 8 to 12 d after estrus (n=102). The proportion of animals with a high number (≥ 2) of large follicles (≥ 7 mm) on Day 0 was lower (P < 0.001) in the 2 GnRHtreated groups than in the control group, while the increase in the number of medium size follicles (4 to 6 mm) on Day 0 was greater (P < 0.02) in the GnRH-puncture group. During superstimulation, the proportion of superovulatory cycles with a high follicular (≥ 10 follicles) response was similar in the control and GnRH-no puncture groups. Within the GnRH-treated animals, follicular and ovulatory responses were greater in the GnRH-puncture than in the GnRH-no puncture group (P < 0.001 to P < 0.02). Despite these changes in follicular and ovulatory responses, however, the mean number of embryos produced did not differ (P > 0.1) among treatments (4.3 ± 0.4, 3.7 ± 0.7, and 5.4 ± 0.8 in control, GnRH-no puncture, and GnRHpuncture groups, respectively). This was due primarily to an increase in the mean numbers of unfertilized ova (P < 0.005) and in degenerated embryos (P < 0.06) in the GnRH-puncture group. Results indicate that the beneficial effects of treatment with GnRH and follicular puncture 2 d prior to superstimulation on follicular and ovulatory responses were limited by an increase in the number of unfertilized ova and degenerated embryos.

Dietary fats varying in their fatty acid composition differentially influence follicular growth in cows fed isoenergetic diets.

The working hypothesis was that dietary fats differing in fatty acid composition would differentially influence ovarian follicular growth. Cows (n = 27) were fed isoenergetic, isonitrogenous, and isofibrous diets containing no added fat (control; CT, n = 7) or diets supplemented with fats containing primarily saturated (SAT, n = 7), polyunsaturated (PU, n = 7), or highly polyunsaturated (HPU, n = 6) fatty acids. Coincident changes in serum lipid metabolites, insulin, and GH and the concentration of IGF-I in large and medium-sized follicles also were examined. Body weights and body condition scores remained similar for all groups throughout the study. Polyunsaturated fat increased (diet x day, P = .06) the number of medium-sized follicles on d 5 through 9 of a synchronized estrous cycle within 3 wk of onset of feeding and maximized (P < .001) this to a fourfold difference at ovariectomy after 7 wk. Fats with predominantly SAT and HPU tended (P < .10) to produce these effects after 7 wk. All fat-supplemented diets increased serum concentrations of total cholesterol (P < .05), GH (P < .05), and follicular fluid IGF-I in large follicles (P < .065) compared to CT but differentially influenced serum concentrations of insulin. Polyunsaturated fat stimulated a marked increase (P < .001) in serum insulin relative to controls within 3 wk, whereas SAT and HPU increased (P < .05) serum insulin only after 6 to 7 wk. We conclude that consumption of PU fatty acids stimulates a greater rate of ovarian follicular growth in cattle compared to CT, AT, and HPU. Future research should investigate the potential role of insulin in mediating PU effects on follicular growth.

Effect of season on follicular dynamics and plasma concentrations of estradiol-17β, progesterone and luteinizing hormone in lactating Holstein cows

Lactating Holstein cows (n=16), averaging 64.1 d in milk, were utilized over 4 replicate months (April, June, August and November) in a shade management system to examine the effects of season on follicular dynamics and plasma concentrations of estradiol (E2), progesterone (P4) and luteinizing hormone (LH). Cows were synchronized to estrus using a combination of Buserelin (GnRH, 8 ug) and prostaglandin F2α (PGF2α, 25 mg) given 7 d apart. Follicular development was monitored daily by ultrasonography, and plasma concentrations of E2, P4 and LH measured by radioimmunoassay. The replicate month had no detectable effects on estrus interval (3.1 ± 0.3 d) or percentage of cows (78.1 ± 9.4%) that expressed estrus following GnRH and PGF2α treatment. Preovulatory follicles grew at faster rates (P<0.01) in June (2.0 ± 0.6 mm/d), than in April (1.1 ± 0.6 mm/d), August (1.0 ± 0.6 mm/d) or November (1.2 ± 0.6 mm/d). First wave dominant follicles were consistently larger in April than in June, August and November. The larger and more persistent size of the first wave dominant follicle in April was associated with an earlier regression of the largest subordinate follicle and a sharper decrease in the number of medium size follicles (6 to 9 mm) by Day 9 of the estrous cycle. Conversely, growth of the first wave dominant follicle was slower and the largest subordinate follicle was more persistent in August than in April, June or November. The proestrous rise in plasma E2 occurred faster (P<0.01) in August (10.1 ± 2.1 pg/d) than in April (4.6 ± 2.1 pg/d), June (5.3 ± 2.1 pg/d) or November (5.9 ± 2.1 pg/d). Concentrations of P4 in plasma increased and reached higher (P<0.01) luteal values in August (15.1 ± 0.6 ng/ml) and November (16.0 ± 0.6 ng/ml) than in April (6.1 ± 0.6 ng/ml) and June (10.6 ± 0.6 ng/ml). There was no detectable effect of month on LH pulse characteristics 48 h post-PGF2α. The maximum size of the corpus luteum (CL) was greatest in November and was related positively to diameter of the ovulatory follicle of the preceding cycle. Results indicated that ovarian follicular development and dominance may be altered during summer months. However, it is uncertain whether these changes can be related to the well-documented low breeding efficiency during warmer months of the year in subtropical environments.

The effect of a GnRH analogue on the dynamics of follicular development and synchronization of estrus in lactating cyclic dairy cows

A GnRH analogue was used to synchronize ovarian follicular development prior to an injection of PGF 2α for the synchronization of estrus in lactating Holstein cows. On Day 12 (estrus = Day 0) of the experimental cycle, cows (n = 8) were injected with 8 μg Buserelin (BUS group), followed by 25 mg PGF 2α 7 d later (Day 19). Control cows (n = 7) received PGF 2α on Day 12 (PGF group). Ovaries were scanned daily via ultrasonography, and plasma progesterone and estradiol concentrations were determined. Sizes of all visible follicles were recorded. Follicles were classified as small (3 to 5 mm), medium (6 to 9 mm), or large (≥ 10 mm). Between Days 12 and 16 of the cycle, the number of large follicles in PGF cows remained unchanged (1.2), whereas in the BUS group, the number of large follicles decreased from 1.3 on Day 12 to 0.5 on Day 15. Only 4 of 7 PGF cows ovulated a second-wave dominant follicle. In the BUS group, 7 of 8 cows ovulated a GnRH analogue induced dominant follicle that was first identified on Day 15. During the follicular phase (last 5 d prior to estrus), plasma progesterone declined in association with CL regression in both groups, and estradiol concentrations increased, reaching higher (P<.0.05) preovulatory peak concentration in BUS cows than in PGF cows (14.0 ± 1.0 vs 10.4 ± 1.1 pg/ml). The number of medium-size follicles was smaller and the number of small-size follicles tended to be higher in BUS cows than in the PGF-treated group. On the day of estrus, the size of the ovulatory follicle (16.1 vs 13.3 mm) and the size difference between the ovulatory and second largest follicle (11.4 vs 6.2 mm) were both larger in BUS cows than in PGF-treated cows, suggesting a more potent dominance effect of the ovulatory follicle in the BUS cows. This study suggests that a GnRH analogue can alter follicular development prior to synchronization of estrus with an injection of PGF 2α in lactating dairy cows.

Factors affecting ovarian compensation after unilateral ovariectomy in gilts.

Two experiments were conducted to identify the temporal limits of an ovarian compensatory response after unilateral ovariectomy (ULO) in gilts and to examine ovarian and hormonal factors that might be related to this response. In the first study, all eight gilts that were unilaterally ovariectomized on d 12, 14 or 16 of an estrous cycle and two of four ULO on d 18 had a significant compensatory increase in ovulation rate per ovary compared with controls. Ovulatory compensation failed to occur, however, in two of four and four of four gilts ULO on d 18 or 19 of an estrous cycle, respectively. In the second study, blood samples were collected from 26 gilts beginning on d 14 of an estrous cycle, and animals were assigned to sham-surgery or ULO on d 17, 18 or 19. Ovarian compensation occurred after ULO on all 3 d of surgery in the second study, but most follicles failed to ovulate and formed large luteinized cysts. The ability to compensate was related positively to the number of medium-sized follicles on the intact ovary at the time of ULO, to an increase in concentrations of follicle-stimulating hormone 12 to 18 h after ULO and to the interval from ULO to the preovulatory surge of luteinizing hormone. In a third experiment, interruption of the ovulatory mechanism and development of cystic follicles similar to those observed in the second study were induced simply by physical manipulation of the d 19 preovulatory ovary while contralateral untouched ovaries ovulated normally.

Serum luteinizing hormone, follicle-stimulating hormone, prolactin and progesterone concentrations and follicular development in the pseudopregnant rat after unilateral ovariectomy.

Serum LH, FSH, prolactin and progesterone concentrations and follicular size were measured in rats during pseudopregnancy after unilateral ovariectomy (ULO) on day 1. This operation did not affect the duration of pseudopregnancy. Following pseudopregnancy compensatory ovulation occurred. After ULO progesterone concentrations remained low as compared with control values but the progesterone secretion per ovary was increased slightly. After ULO, LH concentrations were significantly increased on days 3 and 4 of pseudopregnancy. FSH concentrations were increased 5 h after the operation and again on day 4-7 of pseudopregnancy when compared with sham-operated control animals. During pseudopregnancy prolactin secretion increased each night. Increased prolactin levels were observed 18 and 36 h after ULO. The number of medium-sized follicles had increased 24 h after ULO. From the present study it is concluded that progesterone secretion by the corpora lutea can be increased to a limited degree by the increased release of gonadotrophins. Furthermore, it is concluded that a rapid increase in FSH concentrations after ULO is responsible for the initiation of the compensatory follicular growth.