Antral Follicle Count Group Research Articles

Delayed endometrial preparation for the induction of luteolysis as a potential factor for improved reproductive performance in Angus beef heifers with high antral follicle counts.

Antral Follicle Count (AFC) and anti-Müllerian hormone (AMH) concentrations are reflective for ovarian reserve and have been associated with improved reproductive performance in cattle. Key events for regulation of uterine receptivity are orchestrated by progesterone. As progesterone concentrations are greater in animals with high than low AFC, we tested the hypothesis, if the resulting improved uterine environment will lead to improved conceptus elongation and endometrial response to interferon tau. For four years, 10 heifers with lowest and highest AFC, respectively, were selected from 120 heifers. Reproductive tracts and blood samples for progesterone and AMH analysis were collected after synchronization and insemination. For a recovered conceptus, length was determined, and interferon tau (IFNT) transcript abundance was analyzed. Endometrial transcript abundance of interferon-stimulated gene 15 (ISG15) and oxytocin receptor (OXTR) were analyzed. Progesterone concentrations did not differ between Low and High AFC Group (P= 0.1). A difference in conceptus length was not observed. Endometrial abundance of ISG15 did not differ between Pregnant Low and High AFC heifers. Abundance of OXTR was greater in Open Low AFC than Open High AFC heifers (P< 0.01). Interaction of AMH and Pregnancy Status was determined, with greater AMH in Pregnant than Open High AFC heifers (P< 0.05). Improved uterine environment in High vs. Low AFC heifers did not result in longer conceptuses or improved endometrial response. As the increase in OXTR transcript abundance was only detected in Low AFC heifers, reported differences in reproductive performance might be associated with earlier initiation of luteolysis.

Relationships between antral follicle count and reproductive characteristics of embryo-recipient mares

Mares (n = 77) were evaluated by antral follicle count (AFC) and selected as embryo recipients. Cyclic recipients received embryos between days 4-6 after ovulation. The acyclic recipients received an intramuscular (i.m.) protocol with 5mg of estradiol benzoate (EB) on the day of donor ovulation (D0; D-4 recipient), 3mg of EB on the following day (D1; D-3 recipient), and 3mg of EB (D2; D-2 recipient). Furthermore, 1500mg of progesterone (P4) i.m. given on D0 of the recipient (D4 donor) followed by 1500mg of P4 on the day of ET (D4-6 recipient). On the ET day, the AFC and animals’ weight, body condition score (BCS), corpus luteum diameter, age and degree of uterine edema (UE) were measured. Pregnancy was confirmed on days 12 and 30. Low AFC was defined as ≤11 follicles (n = 43 mares) and high AFC as >11 follicles (n = 34 mares). Data were analyzed by a mixed effect model, including AFC group, reproductive seasonality, and season (P ≤ 0.05). UE was influenced (P = 0.05) by reproductive seasonality. The conception rate was higher (P = 0.016) in recipients with low (79.07 %) than high AFC (61.76 %) and higher (P = 0.005) in cyclic (81.40 %) than anestrus (58.82 %) mares. In addition, we observed a tendency (P = 0.06) for the interaction of AFC*reproductive seasonality, showing that high*anoestrus recipients had the lowest conception rate (37.50 %b) compared to high*cyclic (83.33 %a), low*anoestrus (77.78 %a) and low*cyclic (80 %a). The conception rate was higher in cyclic recipients with low AFC. Furthermore, UE was influenced by reproductive seasonality and mares in anestrus showed a higher degree of UE than cyclic mares.

Effects of maternal antral follicle count in Bos taurus indicus cattle on the genetic merit of male offspring and antral follicle count of female offspring

This study evaluated the effects of antral follicle count (AFC) in female cattle on offspring characteristics. Recently calved multiparous Bos indicus cows (Nelore; n = 222) were evaluated using ultrasonography on random days of their estrous cycle to determine the AFC and were classified into “low” (≤15 follicles), “intermediate” (≥16 and ≤ 29 follicles), and “high” (≥30 follicles) AFC groups. Weight and scrotal circumference (SC) of male offspring from these cows (n = 127) were determined from 20 to 27 months, and the data were added to a genetic evaluation program (economic total genetic merit, MGTe and TOP value) that uses the kinship matrix to evaluate the genetic relationship between animals. The AFC of female offspring from these cows (n = 95) was evaluated to analyze the relationship between the AFC of mothers and daughters. The effects of maternal AFC on the genetic merit of male and female offspring were analyzed using GLIMMIX and GLM, respectively. Correlations were assessed using the Pearson's coefficient. Male offspring of cows with high AFC had superior MGTe (P = 0.005) and TOP values (P = 0.01) than those from cows with low AFC. Additionally, the AFC of mothers was positively correlated with MGTe (R = 0.33; P < 0.0001) and negatively correlated with TOP values (R = −0.32; P < 0.0001). The SC (P = 0.01), but not body weight of the offspring (P = 0.46) was affected by maternal AFC. The daughters' AFC were correlated (R = 0.29; P = 0.004) with mothers' AFC and were influenced by maternal (P = 0.05) but not paternal (P = 0.77) effect. In conclusion, cows with high AFC produced males with greater MGTe, superior TOP values and higher SC. Maternal AFC did not influence the weight of male offspring but was correlated with the AFC of daughters.

Investigation of androgen receptor CAG repeats length in polycystic ovary syndrome diagnosed using the new international evidence-based guideline

BackgroundTo study whether CAG repeat polymorphism of androgen receptor (AR) contributes to the risk of polycystic ovarian morphology (PCOM) with antral follicle count (AFC) ≥ 20 in the context of new international guideline of polycystic ovary syndrome (PCOS).MethodsBlood of 109 PCOS cases and 61 controls were collected for the measurement of AR CAG repeats length by sequencing. The mean number and frequency distribution of CAG repeats length were observed. Detailed analysis was conducted by dividing PCOS cases into low AFC group (L-AFC, AFC < 20) and high AFC group (H-AFC, AFC ≥ 20) according to the new international evidence-based guideline.ResultsThe portion of individuals with lower CAG repeats length in H-AFC group was significantly larger than those with higher CAG repeats length. Logistic model revealed individuals with lower CAG length tended to develop H-AFC.ConclusionLower CAG repeats length in the AR gene of PCOS cases increases risk of PCOM.

Accuracy of ultrasound and visual inspection in antral follicular count in crossbred Holstein cows raised under grazing systems at high altitude.

The objective of this research was to investigate the diagnostic accuracy of post-mortem ultrasound in antral follicle count (AFC) determination and compare it with visual AFC in grazing crossbred Holstein cows, at high altitude in Ecuador. Pre-mortem blood from 80 cows was collected, and AFC and ovarian characteristics were analysed post-mortem by ultrasound and visual techniques. AFC counts were stratified as high, medium or low by terciles. Mean AMH concentration in pre-mortem blood was 280.1 ± 15.53 pg/mL. The AFC obtained by visual inspection (26.9 ± 9.49 follicles) was 23.8% higher than by ultrasound (20.5 ± 7.53 follicles) in all ovaries. Body condition score, age and weight of the cattle did not interact with the count technique. In the low AFC group, visual inspection and ultrasound provided similar AFC results. However, in the Medium- and High-AFC groups, AFC by ultrasound was 14.9% lower than AFC by visual inspection. We confirm that ultrasound can be used with great accuracy for AFC >3 mm (close to the resolution limit) in grazing crossbred Holstein cows at high altitude.

Effects of postpartum diseases on antral follicle count and serum concentration of Anti-Müllerian hormone in dairy cows

The antral follicle count (AFC) and Anti-Müllerian hormone (AMH) concentration are validated markers for ovarian reserve in cattle, but their use as fertility markers is controverse. Here we assessed the effects of postpartum diseases on AFC and AMH concentration, as well as the influence of parity and breed on these parameters. Cows (n = 513, mostly Holstein Friesian and Brown Swiss, parity 3.0 ± 1.8) underwent a single ultrasonography examination 28–56 days after parturition and categorized as having low (n ≤ 15 follicles), intermediate (n = 16–24 follicles), or high (n ≥ 25 follicles) AFC based on objective video analysis of recorded sequences. Blood samples for AMH determination were collected at the time of examination and animals divided into low (< 0.05 ng/ml) and high AMH (≥ 0.05 ng/ml) group, respectively. No effects of postpartum diseases or breed on either AFC or AMH groups could be observed. There was a strong interaction between parity and AFC, primiparous cows having less follicles (13.6 ± 6.2 vs. 17.1 ± 7.0, P < 0.001) than pluriparous cows. The AFC did not affect reproductive parameters or productivity of the cows. In comparison, pluriparous cows with high AMH concentration had shorter calving to first service (86.0 ± 37.6 vs. 97.1 ± 46.7 days, P < 0.05) and calving to conception (123.8 ± 51.9 vs. 135.8 ± 54.4 days, P < 0.05) intervals, but lower milk yield (8440.3 ± 2292.9 vs. 8927.9 ± 2192.5 kg, P < 0.05) compared to cows with low AMH. In conclusion, no effect of postpartum diseases on AFC or AMH concentration of dairy cows could be observed. However, an interaction between parity and AFC, as well as associations of AMH with fertility and productivity in pluriparous cows, were demonstrated.

Low versus high antral follicle count on the fertility of timed AI Nelore heifers

This study aimed to determine the effect of antral follicle count (AFC), and pubertal status on the fertility of beef heifers. In this study, 230 Nelore heifers, 20±2 months of age, were subjected to an estradiol progesterone-based timed artificial insemination (TAI) program. On Day 0 of the TAI protocol, the heifers were examined by transrectal ultrasound to record videos of the ovaries. Later, in the darkroom of the laboratory of images, the videos were analyzed for AFC (≥ 3 mm) of each ovary. Females who failed the first TAI were resynchronized with the same hormonal protocol. The pregnancy status was evaluated by ultrasonography 30 days after each FTAI. The general mean of the AFC was 22.0 follicles. Thus, the heifers were divided into 2 groups according to AFC: Low AFC (˂ 22 follicles, n = 114), and High AFC (≥ 22 follicles, n = 116). No differences (P &gt; 0.05) in the pregnancy per AI (P/AI) were observed between the Low and High AFC groups, and between pubertal and prepubertal categories. The P/AI was not different between heifers that displayed or did not estrus (P = 0.2). However, considering the estrus response of each AFC group, High AFC heifers that displayed estrus had greater P/AI (P = 0.01) than High AFC heifers that did not display estrus. In summary, AFC and pubertalstatus did not affect the fertility of Nelore heifers. In contrast, the P/AI of heifers that did not display estrus was lower than heifers observed in estrus only in the High AFC group.

Anticipation of estrus and ovulation in Nelore heifers with low antral follicle count

This study aimed to establish the relationship between antral follicle count (AFC), the time of puberty, and ovarian response in Nelore heifers subjected to timed-AI (TAI) protocols. Prepubescent Nelore heifers (Bos indicus; n = 30) were examined by transrectal ultrasound and videos of the ovaries were recorded for later CFA (≥ 3 mm). After AFC, heifers were divided into 2 groups, low and high AFC and then further ultrasonic ovarian assessments were performed to determine the onset of puberty. Once puberty was established in all heifers, they were submitted to an FTAI protocol. There were no differences (P &gt; 0.1) between groups on the time of puberty, ovulation rate, the proportion of heifers that expressed estrus, and on preovulatory follicle diameter. However, heifers with low AFC had a smaller ovarian area in the prepubertal period (4.36 ± 0.3 vs. 5.65 ± 0.2 cm2) and, in the FTAI, they had the moment of estrus (48 ± 0.0 vs. 58.5 ± 3.1 h) and earlier ovulation (60 ± 1.8 vs. 72.6 ± 4.7 h) compared to heifers with high AFC (P &lt; 0.05). In addition, heifers in the low AFC group had a higher (P = 0.05) pregnancy per AI (76.9%, 10/13) than the high AFC group (29.4%, 5/17). In this study, heifers with low AFC had the moment of ovulation anticipated and greater pregnancy per AI compared to heifers with high AFC.

PSI-27 Effect of antral follicle count on time intervals to estrus and ovulation in crossbred beef heifers

Abstract While genetics are an unreliable indicator of fertility, antral follicle count (AFC) has recently been recognized as a useful fertility prediction tool in heifers. The aim of this study was to determine if AFC exerts an effect on response to synchronization, time to estrus (TTE), and/or time to ovulation (TTO). Twenty-five crossbred heifers were classified by AFC, either high (&amp;gt; 25 antral follicles), moderate (16–24 antral follicles), or low (&amp;lt; 15 follicles) and utilized in two experiments. In experiment 1, heifers were subjected to the Select Synch + CIDR fixed time artificial insemination (FTAI) protocol. After synchronization, estrus activity was monitored every 3 h until standing estrus was observed. Heifers were examined at 12 and 18 h after observation of standing estrus and every 3 h thereafter to monitor ovulation. In experiment 1, the proportion of heifers that responded to the protocol was not significant (P = 0.71) among classification groups. Time to estrus and time to ovulation were similar between groups (P &amp;gt; 0.90). In experiment 2, heifers were placed with bulls for 60 d and were monitored for signs of estrus every 3 h. Results of experiment 2 indicated that response to estrus synchronization was similar across groups (P = 0.46). Time to estrus was greater (P = 0.04) in the high AFC group than in the low or moderate AFC groups. This may be attributed to lower concentrations of circulating gonadotropins in high AFC heifers, resulting in decreased intrafollicular estradiol. Differences in TTE results between experiment 1 and experiment 2 may be explained by increased reproductive function as heifers continued to mature during the study. These results suggest that differences in TTE may be related to AFC. Additional research is needed to corroborate these results and inform the creation of differing FTAI protocols based on AFC.

Measuring IGF-1 and IGFBP-3 profiles in women seeking assisted reproduction; relationship to ovarian reserve parameters, namely AFC and AMH (study 2).

This is the second of five studies undertaken on women preparing for assisted reproduction, to sequentially examine the relevance of the insulin-like growth factor (IGF) profile (IGF-1, IGFBP-3 and the IGFBP-3/ IGF-1 ratio) which, in children, provides the essential criteria to identify the GH-deficient individual. Whilst our first, published study, focussed on clinical parameters, this study examines its relevance to the two parameters which define the ovarian reserve. The first, that of the antral follicle count (AFC), shows highly significant, sequential changes across 4 age groups ranging from high counts in the younger women &lt;35 years and low counts in the older women, namely those aged 35-39 years, those aged 40-44 years and those aged ≥45 years (p&lt;0.0001). Similarly, the serum levels of anti-Mullerian hormone (AMH), a later introduced marker of ovarian reserve, also showed highly significant sequential changes across the 4 age groups with high levels recorded in the young women and low counts in the older women (p&lt;0.0001). At the higher AFC range, concordance between AFC groups and AMH groups was high at r=0.79 for precise matching and r=0.95 when neighbouring groups were included. The correlation was also clear with inter-quartile AMH levels ranging 27 pmol/L to 50 pmol/L across the higher AFC groups and 8 pmol/L to 10 pmol/L in the lowest groupings. However, IGF profiles showed no significant variations across the entire range, neither for the AFC nor for the AMH groups. Our next study will report on the relevance of the IGF profile to clinical outcomes.

Impact of the newly recommended antral follicle count cutoff for polycystic ovary in adult women with polycystic ovary syndrome.

What is the impact of the newly recommended antral follicle count (AFC) cutoff for polycystic ovary (PCO) on the diagnostic status of polycystic ovary syndrome (PCOS)? Among patients with phenotypes requiring the presence of PCO for diagnosis, approximately half (48.2%) were excluded from having PCOS based on the new AFC cutoff, although these excluded women had worse metabolic and hormonal profiles than the controls and were indistinguishable from the remaining patients with regard to major hormonal and metabolic parameters. In the Rotterdam criteria, PCO is defined as either 12 or more follicles measuring 2-9mm in diameter or an increased ovarian volume >10cm3. Recently, an international PCOS guideline development group recommended an AFC threshold for PCO of ≥20 in adult women when using transducers with a high-resolution frequency, including 8MHz. The current study used a case control design. PCOS was diagnosed according to the Rotterdam criteria. Ultrasonography examinations were conducted with wide band frequency (5-9MHz) transvaginal transducers and the centre frequency was 8MHz. In patients who show both irregular menstruation and hyperandrogenism (HA), a diagnosis of PCOS can be made irrespective of the ovarian criteria change. Patients who were diagnosed according to HA and PCO (n= 86) or irregular menstruation and PCO (n= 443) were initially included among a total of 1390 adult women with PCOS (aged 20-40years). Regardless of the AFC, if the ovarian volume is ≥10cm3, a diagnosis of PCO can still be made. Thus, only patients who had an ovarian volume of <10cm3 were analysed. Subjects who had an AFC of 12-19 and an ovarian volume <10cm3 were designated as the 'low AFC group' (n= 255) and were the main focus of the study because they were excluded from having PCOS based on the new cutoff. Subjects with an AFC ≥20 and an ovarian volume <10cm3 were designated as the 'high AFC group' (n= 101). A total of 562 premenopausal women without PCOS were enrolled as controls. Among patients with irregular menstruation and PCO or HA and PCO phenotypes, approximately half (48.2%, 255/529) were excluded from having PCOS, which corresponded to one-fifth (18.3%, 255/1390) of the total adult patients. However, compared to the control group, these excluded women had worse metabolic profiles and were more androgenised. Notably, they were indistinguishable from the 'high AFC group' with regard to major hormonal and metabolic parameters (BMI and diabetic classification status, and the prevalence of insulin resistance, metabolic syndrome and HA). We cannot exclude the possibility of inter- and intraobserver variation in the evaluation of AFC. With the newly recommended follicle count cutoff, a substantial proportion of women with PCOS might be classified as not having PCOS despite visiting a hospital due to irregular menstruation or hyperandrogenic symptoms. A practical approach to them would involve controlling the menstrual or hyperandrogenic symptoms in hand and regularly evaluating them regarding newly developed or worsening PCOS-related symptoms or metabolic abnormalities. This study was supported by a grant from the Seoul National University Hospital Research Fund (No. 2520140090), Republic of Korea. The authors have no conflicts of interest to disclose. Not applicable.

Comparison of antral follicle count and serum anti Müllerian hormone level for determination of gonadotropin dosing in in-vitro fertilization: randomized trial.

To compare the proportion of women achieving a desired ovarian response following ovarian stimulation when gonadotropin dosing was determined based on antral follicle count (AFC) vs serum anti-Müllerian hormone (AMH) level, in women undergoing in-vitro fertilization (IVF) using the gonadotropin-releasing hormone (GnRH) antagonist protocol. This was a randomized double-blind trial carried out in a university-affiliated assisted reproduction unit. A total of 200 women undergoing their first IVF cycle using the GnRH-antagonist protocol between April 2016 and February 2018 were randomized to determination of gonadotropin dosing based on either AFC or serum AMH level measured in the pretreatment cycle 1 month before the IVF cycle. Patients underwent IVF as per our center's standard protocol. The proportion of subjects achieving a desired ovarian response, defined as retrieval of six to 14 oocytes, was compared between the two study arms. Subgroup analysis of patients with baseline AFC > 5 and those with baseline AFC ≤ 5 was performed. Concordance in AFC and AMH categorization between the pretreatment cycle and the ovarian-stimulation cycle was assessed using Cohen's kappa (κ). There was no significant difference in the proportion of patients achieving a desired ovarian response between the AFC (54%) and AMH (49%) groups (P = 0.479). The median number of oocytes retrieved was nine vs seven (P = 0.070), and the median follicular output rate was 0.54 vs 0.55 (P = 0.764) in the AFC and AMH groups, respectively. Similar findings were observed on subgroup analysis of subjects with AFC ≤ 5 and AFC > 5 at the start of ovarian stimulation (P > 0.05 for all comparisons). There was moderate concordance between AFC and AMH measured in the pretreatment cycle and the stimulation cycle (κ = 0.478 and 0.587, respectively). The proportion of women achieving a desired ovarian response following ovarian stimulation using the GnRH-antagonist protocol is similar when the gonadotropin-dosing algorithm used is based on AFC or serum AMH level. Copyright © 2019 ISUOG. Published by John Wiley & Sons Ltd.

130 Increased antral follicle population at 60 days of gestation in Holstein cows with low antral follicle count

The antral follicle count (AFC) has been pointed out as one of the main factors that influence the efficiency of reproductive biotechniques. Thus, AFC is considered a fertility marker, which has high repeatability in the same animal. The objective of this study was to evaluate AFC in cows at AI (nonpregnant) and at two moments of gestation (30 and 60 days). For this, 35 pregnant cows, aged 23 to 99 months, confined in a freestall system, kept in the same environmental conditions (24°46'28”S, 49°56'42”W) and under the same feeding conditions, were selected for this study. The same technician performed the AI of the females, after detection of natural or induced oestrus, with 25mg of Dinoprost (Lutalyse) intramuscularly (IM). We established AFC (antral follicles ≥3mm diameter) using a convex intravaginal transducer at the time of AI and at 30 and 60 days of pregnancy. Females were distributed in groups of low (≤18 follicles, n=11), intermediate (≥24 and ≤28 follicles, n=11), or high AFC (≥30 follicles, n=13). Data were analysed using a generalized linear mixed model (Minitab version 18.1), with significance declared at P ≤ 0.05. In the low AFC group, there was an increase in the mean number of antral follicles in relation to gestation, presenting 14.82±1.36 at the time of AI and 23.45±2.31 and 35.18±3.17 at 30 and 60 days of pregnancy, respectively (P&amp;lt;0.0001). Similarly, the intermediate group presented an increase in mean AFC, from 24.64±0.76 at the time of AI and 29.27±3.32 at 30 days to 43.36±5.91 at 60 days of gestation (P=0.001). There was no difference in the high AFC group at different moments, as these presented 41.92±3.50 at the time of AI, 38.31±3.96 at 30 days, and 50.92±5.49 at 60 days of pregnancy. Although mean AFCs differed (P&amp;lt;0.0001) between the low, intermediate, and high AFC groups at time of AI and at 30 days of gestation, there were no differences in AFC between the three groups at 60 days of gestation (P=0.329). In conclusion, low AFC females presented an increase in the number of antral follicles at gestation moments 30 and 60 days from AI. This finding reinforces the strategy of using this category more efficiently for ovum pickup at the beginning of gestation.

Relationships between the antral follicle count, steroidogenesis, and secretion of follicle-stimulating hormone and anti-M\xfcllerian hormone during follicular growth in cattle

BackgroundThe antral follicle count (AFC) in mammalian ovaries positively correlates with female fertility. To clarify the causes of differences in fertility between low and high AFC cows, we investigated follicular growth dynamics and hormone concentrations in plasma, follicular fluid, and in vitro growth (IVG) media at different stages of follicular growth.MethodsSeven cows were divided into high AFC (n = 4, > 30 follicles) and low AFC (n = 3, < 30 follicles) groups based on the peak AFC detected by ultrasonography. These cows were subjected to estrous synchronization, daily ovarian ultrasonography, and blood collection. Their follicular fluid was collected from dominant follicles at different stages (selection, luteal, and ovulatory phases). In another experiment, we cultured oocyte-cumulus-granulosa cell complexes collected from early antral follicles (< 1 mm) for 12 days. Estradiol-17β (E2), testosterone (T), progesterone (P4), and anti-Müllerian hormone (AMH) concentrations in follicular fluids and plasma were measured. Plasma follicle-stimulating hormone (FSH) concentrations were examined. E2, P4, and AMH concentrations were also measured in IVG media.ResultsThe numbers of small (< 4 mm) and intermediate (4–8 mm) follicles were larger in the high AFC group than in the low AFC group (P < 0.05). The number of intermediate follicles was stable in the low AFC group, indicating consistent development. However, the number of these follicles fluctuated in the high AFC group. Plasma FSH concentrations were higher, whereas E2 and T concentrations were lower in the low AFC group (P < 0.05). E2 concentrations and the E2/P4 ratio in ovulatory follicles and IVG media on day 8 were higher in the high AFC group (P < 0.05). AMH concentrations in plasma and IVG media (P < 0.01) were higher in the high AFC group.ConclusionsThe weaker response to FSH of granulosa cells caused low E2 production in the low AFC group, resulting in high FSH concentrations and the consistent development of intermediate follicles. Conversely, higher E2 concentrations suppressed FSH secretion in the high AFC group. Granulosa cells in the high AFC group had the ability to produce more AMH than those in the low AFC group throughout IVG culture.

Is corifollitropin alfa effective in controlled ovarian stimulation among all poor ovarian responders? A retrospective comparative study

Several studies have compared the effectiveness of corifollitropin alfa versus daily gonadotropins in poor ovarian responders (PORs) undergoing controlled ovarian stimulation (COS), showing conflicting results in terms of IVF outcomes. Given the heterogeneity of patients included in the classification of POR according to ‘Bologna criteria’, the aim of this study was to evaluate the impact of corifollitropin alfa in two different categories of POR distinguished according to patients’ antral follicle count (AFC). We retrospectively evaluated 104 infertile POR, split into two groups according to AFC (Group A ≤ 5; Group B > 5) and subgroups according to the ovarian stimulation regimen (corifollitropin alfa plus daily gonadotropins (Subgroup 1) versus daily gonadotropins alone (Subgroup 2)). Outcome measures were total oocytes, MII oocytes, total embryos, follicular output rate (FORT), implantation rate (IR), clinical pregnancy rate (CPR), miscarriage rate (MR), and live birth rate (LBR). Subgroup A1 experienced a lower number of total oocytes, MII oocytes, total embryos, and FORT (p < .05) in comparison to Subgroup A2, while no difference was found when comparing Subgroups B1 and B2. No difference was found between subgroups even in terms of IR, CPR, MR, and LBR. In conclusion, corifollitropin alfa may be as effective as daily gonadotropins in POR with AFC > 5 undergoing COS, while it might be inferior to daily gonadotropins in POR with AFC ≤ 5.

Relationship between the antral follicle count in bovine ovaries from a local abattoir and steroidogenesis of granulosa cells cultured as oocyte-cumulus-granulosa complexes.

The antral follicle count (AFC) is used as an indicator of cow fertility. We herein investigated the relationship between AFC and the steroidogenesis of granulosa cells and confirmed the developmental competence of oocytes derived from early antral follicles (0.5–1.0 mm) using in vitro growth culture. Slaughterhouse-derived ovaries were divided into high (≥ 25) and low (< 25) AFC groups based on AFC (≥ 2.0 mm). Oocyte-cumulus-granulosa complexes (OCGCs) collected from early antral follicles were cultured for 12 days. The total number, viability, and diameter of granulosa cells and estradiol-17β and progesterone production during the culture were evaluated. Surviving oocytes on day 12 were subjected to in vitro maturation, and their volume and nuclear status were evaluated. Some oocytes were subjected to the evaluation of developmental competence to blastocysts. Although the total number and viability of granulosa cells did not differ between the groups, granulosa cell diameters were smaller in the high AFC group than in the low AFC group. The estradiol-17β and progesterone ratio on day 8 was higher in the high AFC group than in the low AFC group. Oocyte volumes and nuclear maturation rates were greater in the high AFC group than in the low AFC group. The development rate to blastocysts was 9.1% in the high AFC group, while no oocytes developed to blastocysts in the low AFC group. Therefore, estradiol-17β production by granulosa cells appears to be greater in high AFC cattle than in low AFC cattle, thereby promoting the acquisition of oocyte competence.

Ovarian antral follicle populations and embryo production in cattle

Reproductive biotechniques such as embryo production are important tools to increase the reproductive performance in cattle in a short time. In this context, the antral follicle count (AFC), which reflects the population of antral follicles present in an ovary, has been indicated as an important phenotypic characteristic related to female fertility and closely correlated to the performance of in vivo and in vitro embryo production (IVEP). A positive correlation was evidenced between AFC and oocyte retrieval by ovum pick up (OPU) sessions and and with the number of embryos produced. Several studies have reported that females with a high AFC had greater embryo yields compared to those with medium and low AFC. However, controversial results were obtained by studies conducted in different bovine breeds. Many conflicting data may be due to the differences in the experimental design, particularly regarding the classification of animals in AFC groups, subspecies particularities, herd aptitude or even issues related to animal management. Therefore, aspects such as the choice of donor, type of aspirated follicles and the stage of follicular wave need to be clarified. Thus, this text aims to discuss the use of AFC as a reproductive tool and its applications in the in vivo and in vitro production of embryos, besides describing consistent results and new challenges regarding AFC and embryo production.

144 EFFECT OF ANTRAL FOLLICLE COUNT IN BEEF HEIFERS ON IN VITRO FERTILIZATION/PRODUCTION

Our objective has been to compare the IVF and in vitro production (IVP) of embryos from low and high antral follicle count (AFC) heifers. This is the fourth year of the study with years 1 to 3 reported individually. For this report, we add data for the fourth year and present a combined analysis (years 1 to 4) for the first time. Each year, AFC was determined on ~120 Angus heifers using transrectal ultrasonography. Ten heifers with the lowest AFC and 10 heifers with the highest AFC and all with evidence of oestrous cyclicity were synchronized with two 5-mL injections of PGF2α 11 days apart. Half were harvested on Day 5 to 6 and half on Day 15 to 16 of the oestrous cycle. The IVF procedure was slightly modified each year. For year 4, the IVF procedure included protocols for semi-defined media and was as described (IVP Protocol, P. J. Hansen’s Laboratory, University of Florida). Cumulus-oocyte complexes (COC) from follicles less than 8 mm in diameter were cultured in maturation medium (5% CO2; 38.5°C) for 24 h. Matured COC were fertilized using thawed frozen semen from a bull that was purified using isolate. Motile spermatozoa were added to COC in fertilization medium at a final concentration of 1 × 106 spermatozoa per mL. About 24 h later, presumptive zygotes were placed in micro drops of development medium under oil, and cultured (5% CO2; 5% O2; balance N2; 38.5°C). On Day 3 and 8 after fertilization, cleavage and blastocyst development rates, respectively, were assessed. Data were analysed using the MIXED procedure of SAS (SAS Institute Inc., Cary, NC, USA) and the model included the effects of year (1 to 4), group (high or low AFC), and their interaction. The year × group interaction was not significant (P &gt; 0.10). Low AFC heifers, compared with high AFC heifers, had fewer numbers of COC (P &lt; 0.0001; 12.8 ± 1.83 v. 31.9 ± 1.86), fewer numbers of COC that cleaved (P &lt; 0.0001; 8.0 ± 1.38 v. 21.6 ± 1.40), and fewer numbers of COC that developed to the blastocyst stage (P &lt; 0.0001; 1.7 ± 0.58 v. 5.7 ± 0.58). Year affected the numbers of COC that cleaved (P &lt; 0.003) and the numbers of COC that developed to the blastocyst stage (P &lt; 0.0001). Year also influenced the percentage of COC that cleaved (P &lt; 0.0002) and the percentage of COC that developed to blastocysts (P &lt; 0.0001). Group (AFC) did not influence (P &gt; 0.19) the percentage of COC that cleaved (61.2 ± 2.83 v. 66.4 ± 2.83%, for low v. high AFC, respectively). Low AFC heifers had a lower (P &lt; 0.002) percentage of COC that developed to blastocysts (10.3 ± 1.52%) than high AFC heifers (17.6 ± 1.52%). These results indicate that high AFC heifers, compared to low AFC heifers, have more COC recovered, more COC cleaved, and more COC developed to the blastocyst stage. The percentage of COC cleaved did not differ between AFC groups; however, the percentage of COC that developed to the blastocyst stage was greater for high than low AFC heifers. This suggests a potential advantage in maternal to embryonic transition for high compared with low AFC heifers.

New automated AntiMüllerian hormone (AMH) assays are more reliable than the manual assay in patients with reduced antral follicle count (AFC)

To improve quality of AMH measurements, fully automated AMH assays have been recently developed. Yet, analytical data available failed to give emphasis on the necessary relationship between AMH and the number of AMH-producing antral follicles as a quality reference. Moreover, the relative performance of new automated and manual AMH assays in subgroups of patients displaying low, intermediate, and high AFC remained undetermined. Therefore, we decided to investigate the reliability and concordance of AMH levels obtained by one manual and two automated AMH assays in different AFC populations. Prospective study. Serum AMH levels were measured in frozen-thawed serum aliquots from 211 women, aged 24 to 43 years, using one manual (AMH Gen II) and two fully automated (Access AMH and Elecsys AMH) assays. AFC was performed on cycle-days 1-5 (same day of bloodwork) under strictly standardized conditions and sorted into 3 groups (tercile values): low AFC (3-12 follicles; n=73), intermediate AFC (13-20 follicles; n=65), and high AFC (21-84 follicles; n=73). The strenght of relationships between AFC and AMH levels was assessed by the Spearman correlation test and Spearman correlation coefficients were compared using the Fisher r-to-z transformation. Concordance among AMH assays was assessed graphically using the Bland-Altman plots and by Passing-Bablok regression. Overall, circulating AMH levels were lower (P< 0.001) with Access AMH (-16%) and Elecsys AMH (-20%) than with AMH Gen II. The limited amount of AMH values situated outside the ± 2 standard deviation interval in Bland-Altman plots (4.7% between AMH Gen II and Access AMH; 4.3% between AMH Gen II and Elecsys AMH; and 2.8% between Access and Elecsys AMH) indicated adequate concordance among assays, a phenomenon that tended to be improved in the low AFC group. Further, AFC was strongly correlated with AMH levels obtained with the 3 assays (r=0.83, P<0.001; r=0.83, P<0.001; and r=0.83, P<0.001, respectively) thereby confirming their comparable reliability. It is noteworthy that, in the low AFC group, AMH levels obtained by Access AMH and Elecsys AMH showed a stronger correlation with AFC (r=0.63; P<0.03 and r=0.65; P<0.005, respectively) than the AMH Gen II assay (r=0.52). This improved performance was not observed in the intermediate and the high AFC groups. As compared to conventional AMH Gen II assay results: 1. Serum AMH concentrations were -16% and -20% lower with Access AMH and Elecsys AMH, respectively, but concordance among techniques was adequate; 2. It was remarkable that both automated AMH assays were significantly more strongly correlated with AFC than the manual assay in patients endowed with few antral follicles. This constitutes a considerable advantage of the new AMH assays because this subset of patients is the main target for accurate ovarian function assessments.

Assessment of ovarian reserve: Anti-Mullerian hormone versus follicle stimulating hormone.

Background:This study aimed to evaluate the strength of anti-Mullerian hormone (AMH) and follicle stimulating hormone (FSH) in reflecting the antral follicle count (AFC) in infertile females.Materials and Methods:This cross-sectional study was conducted on 160 females, visiting infertility clinic for assisted reproduction. Serum samples collected on the 3rd day of the cycle were assayed for FSH, luteinizing hormone, and AMH while AFC was assessed via transvaginal ultrasound. The study cohort was segregated into three groups based on AFC.Results:Chronological age and FSH was significantly high in females with very low AFC (P < 0.01 and 0.009, respectively), yet they failed to discriminate patients with normal and higher follicle count (P = 0.65 and 0.84). Conversely, AMH reported highly significant difference between very low AFC and with those having either normal AFC (P = 0.002) or higher AFC (P = 0.001). Moreover, a significant difference in AMH was observed between normal and higher AFC group (P = 0.04).Conclusion:Compared to female’s age and FSH, AMH is superior in clustering study cohort on the bases of antral follicular pool, especially in setups with nonavailability of technological expertise to assess AFC. Incorporation of AMH along with other biomarkers improves estimation of baseline ovarian reserve, required to standardize dose for optimum response; avoiding the risk of failure to retrieve oocyte or inappropriate stimulation leading to ovarian hyperstimulation syndrome. Further prospective studies are required to ascertain its role in predicting the outcomes of ART in such patients.