121 Anti-Müllerian hormone cutoff values for the selection of oocyte donors in the Gir (Bos indicus) breed

2021 ◽  
Vol 33 (2) ◽  
pp. 168
Author(s):  
L. F. Feres ◽  
L. G. B. Siqueira ◽  
L F. M. Pfeifer ◽  
L. L. Santos ◽  
J. H. M. Viana

Anti-Müllerian hormone (AMH) has a well-known association with antral follicle count (AFC) and the number of oocytes recovered by OPU in cattle. However, to be used as a criterion to select candidate donors for ART, AMH reference values must be established for each breed. The aim of this study was to estimate AMH cutoff values for selecting oocyte donors in Gir (Bos indicus) cattle. A blood sample was collected at a random day of oestrous cycle from Gir heifers (n=120) at 23.3±0.5 months of age, and the plasma was stored for further AMH analysis by ELISA, using a commercial kit (AL114, AnshLabs) in a private laboratory (LEAC, Sao Paulo, Brazil). Data from 506 ovum pickup-invitro embryo production (OPU-IVEP) sessions (4.2±0.2 per donor) performed in these heifers from 2017 to 2019, starting 120 days after blood sampling, were then analysed. Donors were ranked in quartiles according to the number of total oocytes recovered, viable oocytes recovered, and embryos produced. Those classified in the first quartile for each endpoint were considered poor responders. The AMH values were then analysed using the receiver operating characteristic (ROC) curve, and cutoff values for each endpoint were estimated based on best values for sensitivity and specificity. A second analysis was performed to check which cutoff value would result in a significant increase in average for each endpoint. The OPU-IVEP data were ranked according to corresponding AMH values (smaller to greater), and the average of the selected group of donors was compared with the original group by ANOVA, using the PROC GLM of SAS (SAS Institute Inc.), after successive exclusion of donors with lower AMH concentrations. Results are shown as mean±s.e.m. On average, heifers presented AMH concentrations of 932.9±48.4pgmL−1 (ranging from 112.7 to 2044.8pgmL−1) and produced 32.3±1.5 total oocytes, 25.9±1.4 viable oocytes, and 5.3±0.4 embryos per OPU-IVEP session. The heifers ranked in the third and last quartiles yielded more total oocytes, viable oocytes, and blastocysts than those ranked in the first quartile (P<0.05). Based on the total number of oocytes recovered (59.2±3.1), heifers of the last quartile had an estimated AFC >60. The area under the ROC curve (AUC) for total oocytes, viable oocytes, and embryos produced were 0.77 (P<0.0001), 0.75 (P<0.0001), and 0.72 (P=0.0003), and the estimated cutoffs for AMH concentration were 761.4, 622.5, and 681.5pgmL−1, respectively. After successive exclusions of donors with lower AMH values, averages were greater (P<0.05) at a cutoff value of 632.4pgmL−1 for total oocytes (36.9±1.6 vs. 32.3±1.5; +15.3%), 672.3pgmL−1 for viable oocytes (30.3±1.4 vs. 25.9±1.4; +19.4%), and 776.8pgmL−1 for embryos produced (6.6±0.3 vs. 5.3±0.4; +23.4%). However, the use of these cutoff values to increase the averages would result in the exclusion of 32.8, 37.9, and 50.0% of the potential donors for each endpoint, respectively. In summary, these results demonstrated that an AMH concentration of approximately 700pgmL−1 can be used to select donors with higher oocyte yield and thus to increase IVEP outcomes in Gir heifers. This research was supported by Fazendas do Basa, Fapemig CVZ APQ 03430-17.

2019 ◽  
Vol 31 (1) ◽  
pp. 10 ◽  
Author(s):  
M. M. Seneda ◽  
F. Morotti ◽  
A. F. Zangirolamo ◽  
N. C. da Silva ◽  
T. K. Sanches ◽  
...  

The antral follicle count (AFC) is an important tool in the selection of bovine females destined for biotechnology. However, little is known about AFC in prepubertal and pubertal heifers. Some challenges inherent to the physiology of young females must be considered to achieve efficient rates with different procedures, such as ovum pick-up and IVF. This paper covers some important topics about ovarian physiology related to the population of antral follicles and reproductive efficiency in young female cattle.


2018 ◽  
Vol 30 (1) ◽  
pp. 143
Author(s):  
F. Morotti ◽  
R. Moretti ◽  
G. M. Gomes dos Santos ◽  
K. C. Silva-Santos ◽  
M. M. Seneda

Two experiments in Nelore cows subjected to conventional protocol for timed AI (TAI) were designed to compare the influence of antral follicle count (AFC; 1) on the ovarian follicular dynamics and (2) conception rates. First, multiparous cows with high (G-high, ≥45 follicles; n = 43) and low AFC (G-low, ≤15 follicles; n = 32) were selected to receive a conventional oestradiol/progesterone-based TAI protocol for monitoring ovarian follicular dynamics. Second, AFC from 962 multiparous suckling cows (40 ± 15 days postpartum) subjected to a conventional protocol for TAI were classified as G-high (≥45 follicles; n = 194), G-intermediate (≥20 and ≤40 follicles, n = 397), and G-low AFC (≤ 15 follicles; n = 243). Pregnancy status was evaluated 30 to 35 days later. Data were analysed using ANOVA followed by the Tukey test and model of logistic regression (P ≤ 0.05). In the first study, the AFC were greater (P ≤ 0.05) in G-high than in G-low, but remained stable in both groups during the evaluations. The ovarian diameter, perimeter, and area were greater (P ≤ 0.05) in G-high than in G-low (28.3 ± 3.9 v. 20.5 ± 3.2 mm, 100.9 ± 13.2 v. 73.8 ± 13.9 mm, and 67.5 ± 16.4 v. 37.7 ± 11.7 mm2; respectively). Follicular growth rate, time of ovulation, corpus luteum diameter, and progesterone concentration were similar (P > 0.05) for cows with low or high AFC. Dominant follicles showed greater (P ≤ 0.05) diameter in G-low than G-high at D4 (7.3 ± 2.2 v. 6.2 ± 1.4 mm; P = 0.069, tendency), D8 (11.2 ± 1.8v. 9.5 ± 1.8 mm), D9 (12.3 ± 1.7 v. 10.6 ± 1.7 mm), and D10 (13.4 ± 1.3 v. 12.2 ± 1.8 mm), and the estimated diameter of the ovulatory follicle tended (P = 0.08) to be higher in G-low than in G-high (14.4 ± 1.5 v. 13.4 ± 2.1 mm). In the second study, AFC of G-low cows showed higher (P ≤ 0.05) conception rate (61.7%) than those of G-intermediate (52.9%) and G-high (49.5%) cows. In Nelore cows subjected to ovulation synchronization protocol, cows with high AFC showed higher ovarian measurements, but those with low AFC had large follicular diameters. Additionally, the cows with low AFC showed higher conception rate than groups with intermediate and high counts.


2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Marina Amaro de Lima ◽  
Fábio Morotti ◽  
Bernardo Marcozzi Bayeux ◽  
Rômulo Germano de Rezende ◽  
Ramon Cesar Botigelli ◽  
...  

Abstract We evaluated the effect of the antral follicle count (AFC) on ovarian follicular dynamics, pregnancy rates, progesterone concentrations, and transcriptional patterns of genes in Nelore cattle (Bos taurus indicus) after a timed artificial insemination (TAI) programme. Cows were separated based on the AFC, and those with a high AFC showed a larger (P < 0.0001) ovarian diameter and area than those with a very low AFC. Females with a very low AFC exhibited a larger (P < 0.01) diameter of the dominant follicle at TAI (13.6 ± 0.3 vs. 12.2 ± 0.4 mm) and a tendency (P = 0.06) to have different serum progesterone concentrations (2.9 ± 0.3 vs. 2.1 ± 0.3 ng/mL; on day 18, considering day 0 as the beginning of the synchronization protocol) than those with a high AFC. The pregnancy rate was higher (P ≤ 0.05) in animals with a very low (57.9%) and low (53.1%) AFC than in those with a high AFC (45.2%). The expression of genes related to intercellular communication, meiotic control, epigenetic modulation, cell division, follicular growth, cell maintenance, steroidogenesis and cellular stress response was assessed on day 5. In females with a low AFC, 8 and 21 genes in oocytes and cumulus cells, respectively, were upregulated (P < 0.05), while 3 and 6 genes in oocytes and cumulus cells, respectively, were downregulated. The results described here will help elucidate the differences in ovarian physiology and the reproductive success of Bos indicus females with a low or high AFC.


Introduction: Poor ovarian responders are the most challenging patients in reproductive medicine and no successful treatment has been proposed. Androgens are thought to play an important role during early folliculogenesis and diminished levels are associated with decreased ovarian sensitivity to follicle-stimulating hormone. This study aimed to determine whether pretreatment with testosterone improves the results in poor responders undergoing in vitro fertilisation (IVF). Materials and methods: This observational pilot study enrolled 33 poor responders undergoing IVF. Eleven patients were pretreated with 250 mg intramuscular testosterone and compared to a control group of 22 patients. The participants were tested for free testosterone, dehydroepiandrosterone sulfate, sex hormone binding globulin, and anti-mullerian hormone (AMH). Results: The two groups had similar baseline characteristics. Significant improvement was reached in the hormones free testosterone, dehydroepiandrosterone sulfate, and sex hormone binding globulin in the testosterone-pretreatment group. No difference was detected in antral follicle count (5.06 versus 4.24); AMH (0.51 versus 0.53), mature oocytes (2.2 versus 2.32), and the number of embryos (1.2 versus 1.33) between the study and control groups, respectively. There was a slow improvement in fertilisation rate but without any significance (62.97% versus 57.61%). However, the cancellation rate of the ovarian stimulation was much greater in the control group (18.18%) in comparison with the study group (0.0%). Pregnancy rate (PR) in the testosterone group was higher than controls (PR per cycle: 27.3% versus 4.6; p=0.09). Conclusion: Based on the limited number of patients studied, pretreatment with testosterone seems to improve PR and cancellation rate in poor responders but failed to affect antral follicle count, AMH, and the number of mature oocytes and embryos. Given these results, further research would provide more certainty.


2019 ◽  
Vol 136 ◽  
pp. 10-14 ◽  
Author(s):  
Fábio Lucas Zito de Moraes ◽  
Fábio Morotti ◽  
Camila Bortoliero Costa ◽  
Paula Alvares Lunardelli ◽  
Marcelo Marcondes Seneda

2020 ◽  
pp. 2171-2178
Author(s):  
Sheila Merlo Garcia ◽  
Paula Alvares Lunardelli ◽  
Kleber Luciano Ancioto ◽  
Eduardo Cardoso de Oliveira ◽  
Larissa Zamparone Bergamo ◽  
...  

This study aimed to evaluate the potential of Girolando (Bos taurus × Bos indicus) cows with high and low antral follicle counts (AFC) for the in vitro production of bovine embryos (IVEP), as well as the pregnancy rates of the recipients of these embryos. Girolando cows (Bos taurus × Bos indicus) were classified as high and low AFC when they had 35-52 (n = 13) and 11-17 follicles (n = 15), respectively. All animals were subjected to repeated follicular aspiration [Ovum pick-up (OPU)] and subsequent IVEP sessions. The synchronization protocol of the recipients was performed on a random day of the estrous cycle (Day 0) with the implantation of progesterone, estradiol benzoate, and prostaglandin. The high AFC group had higher aspirated oocyte/OPU (42.6 ± 5.2 vs. 14.6 ± 1.9; p < 0.01) and cultured oocyte/OPU (38.1 ± 6.6 vs. 12.3 ± 2.8; p < 0.01) averages as well as a higher blastocyst percentage on D7 (23.0 ± 1.0% vs. 18.4 ± 1.5%; p < 0.05) and higher pregnancy rate (42.7 ± 2.7% vs. 39.7 ± 4.6%; p < 0.05) than the low AFC group. Thus, we can conclude that animals with high AFC had better IVEP success rates than animals with low AFC.


2020 ◽  
Vol 32 (4) ◽  
pp. 383 ◽  
Author(s):  
Hayley McGrice ◽  
Jennifer M. Kelly ◽  
David O. Kleemann ◽  
Karen L. Kind ◽  
Alissa J. Hampton ◽  
...  

The use of juvenile invitro embryo transfer (JIVET) is limited by variation between prepubertal lambs in ovarian response to exogenous gonadotrophins. In cattle, anti-Müllerian hormone (AMH) is a predictive endocrine marker of antral follicle count. In this study we measured plasma AMH concentrations in lambs at 3 and 5 weeks of age and determined associations between AMH concentrations and ovarian response to gonadotrophins and invitro blastocyst production at 6–8 weeks of age in a JIVET program. At 5 weeks, AMH (n=38) was positively correlated with surface antral follicle count (r=0.87, P&lt;0.001), blastocysts produced (r=0.92, P&lt;0.001) and blastocysts produced as a proportion of oocytes collected (r=0.44, P&lt;0.01) or cleaved (r=0.43, P&lt;0.01). Similar associations were observed between AMH at 3 weeks (n=30) and follicle number (r=0.70, P&lt;0.05) and blastocysts produced (r=0.87, P&lt;0.05). Lambs with high (&gt;2.2ngmL−1) compared with medium (0.4–2.2ngmL−1) and low (&lt;0.4ngmL−1) AMH at 5 weeks had more antral follicles (mean (±s.e.m.) 118.7±13.9 vs 68.2±8.1 and 30.4±12.3 respectively; P&lt;0.05) and more blastocysts produced (mean (±s.e.m.) 54.9±6.9 vs 18.9±4.0 and 7.5±6.1 respectively; P&lt;0.05). These results suggest that AMH concentration at 5 weeks of age can be used to select donor lambs which enhance the success of JIVET programs.


2016 ◽  
Vol 28 (2) ◽  
pp. 196
Author(s):  
R. Maculan ◽  
R. G. Rosa ◽  
R. R. Bonfim ◽  
G. L. Vasconcelos ◽  
T. L. C. Pinto ◽  
...  

The intense selection of characteristics related to animal production may affect negatively the reproductive efficiency of beef cattle. Thus, the search for characteristics that indicate fertility is readily justifiable in the selection of bovine females. The aim was to evaluate possible associations between the antral follicle counts (AFC), external genitalia measurements, and reproduction efficiency in Tabapuã (a Zebu breed from Brazil) females. The AFC was evaluated in Nulliparous heifers (n = 162) and cows (n = 429) by directly counting all follicles ≥3 mm in diameter with ultrasound. From the frequency distribution of the AFC, animals were divided into groups of high (>50 follicles), average (28–50 follicles), and low (<28 follicles), according to Burns et al. (2005). The vulva width was determined by measuring the distance between the lateral borders of the vulva with a digital caliper placed at a 90-degree angle from the half point of the rima vulvae. Two official reproductive efficiency indexes, adopted by the Brazilian Zebu Breeders Association (ABCZ), were used: age at first calving and calving interval. All statistical analyses were performed using SAS (SAS Institute Inc., Cary, NC, USA). An initial complete model was submitted to logistic regression and all body measurements, such as weight, rump height, and width at pins and hooks, were later excluded because they had no relationships with AFC. The AFC class and parity effects of vulva width were submitted to the PROCGENMOD procedure and the reproductive efficiency indexes were regressed onto age, vulva width and the AFC using the PROCREG procedure. Vulva width was greater (P = 0.05) in the high-AFC class cows (8.81 ± 0.12 cm) in comparison to those of the low (8.38 ± 0.13 cm) and average (8.42 ± 0.11 cm) classes, and was not influenced (P = 0.08) by parity (8.37 ± 0.11 and 8.77 ± 0.14 cm for heifers and cows, respectively). Vulva width was larger in cows with lower calving intervals (r2 = –0.21 P = 0.0008), but was not correlated with age at first calving (P = 0.78). Antral follicle counts were similar (P = 0.71) between heifers (38.6 ± 23.96) and cows (38.9 ± 28.00). Calving intervals were shorter in females with higher AFC (r2 = –0.17 P = 0.0064). Age was slightly related to a decrease in the AFC (r2 = –0.066 P < 0.0001). Vulva width and antral follicle count were correlated and were indicators of reproductive efficiency in females of the Tabapuã breed under the present experimental conditions. To our knowledge, this is the first report where vulva width, a fairly simple to detectable phenotype, was correlated with fertility.


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