Search

A feasible and non‐invasive luteal function assessment which enables timely intervention to improve progesterone (P4) support at early pregnancy is not well‐established. The aim of this study was to determine the correlation among morphological and functional assessment methods of corpus luteum (CL) on Day 5 (D5) following timed‐artificial insemination effect on luteal blood perfusion (LBP), CL diameter and serum P4 concentration. Beef heifers (n = 89) were synchronized and inseminated (D0). On D5, CL was scanned by colour‐Doppler ultrasonography using an LBP score (0 = absent; 3 = high blood perfusion); CL diameter was obtained and blood was collected. Diameter of CL had a positive linear effect on P4 concentration (p <.001); and larger CL diameter increased the probability to become pregnant (p <.05; odds ratio =1.21). Heifers with a CL larger than 14.95 mm had a higher pregnancy rate than heifers with a 14.95 mm or smaller CL (p <.05). Animals with LBP 2 and 3 had larger CL when compared to scores 0 and 1 (p <.001). Scores 1, 2 and 3 can accurately estimate heifers with higher P4 (accuracy =0.79, 0.72 and 0.61 respectively). In conclusion, LBP on Day 5 post‐TAI allows to estimate heifers with higher P4 and larger CL size, and a larger CL diameter was positively associated with pregnancy rate. [ABSTRACT FROM AUTHOR]

High‐density lipoprotein (HDL) is the main lipoprotein in the follicular fluid, and it has anti‐inflammatory, antioxidant and cryoprotectant properties. The anti‐inflammatory potential and antioxidant potential are derived from its lipid composition, especially the apolipoprotein AI (ApoAI) and paraoxonase 1 (PON1). The aim of this study was to evaluate the effect of HDL during in vitro maturation (IVM) on oocyte maturation and early bovine embryo development. For this, cumulus–oocyte complexes (COCs) were obtained from bovine ovaries collected at a local slaughterhouse. COCs (n = 2,250) were allocated into three groups (n = 50 COCs/group) according to the addition of HDL protein (HDL‐P) during IVM for 22 hr: 0 (control), 50 and 150 mg/dl. After IVM, COCs were inseminated (in vitro fertilization) and cultivated for 7 days. Total cholesterol concentration, total protein, triglycerides and ApoAI concentrations on IVM medium increased proportionally to HDL‐P addition. However, PON1 activity was not detected in any treatment. The addition of HDL‐P did not affect nuclear maturation rate, endogenous reactive oxygen species and glutathione levels in COCs (p > 0.05). The highest HDL‐P concentration (150 mg/dl) decreased cleavage and blastocyst rate (p < 0.05). Moreover, the HDL‐P 150 mg/dl group had lower cellular count/blastocyst than the 50 mg/dl group (p < 0.05). However, the addition of HDL‐P did not affect relative gene expression of evaluated genes. In conclusion, the complex HDL/ApoAI obtained from human plasma, in the absence of PON1 activity during in vitro oocyte maturation, decreased initial embryo development. [ABSTRACT FROM AUTHOR]

Background Estradiol (E2) receptors mediate E2 effects on cell proliferation and apoptosis under normal and pathological conditions. However, the mechanisms involved in E2 signaling are not, completely understood. The objectives in this study were to evaluate the expression of estrogen receptors (ESRs) during follicular selection in cattle, and the effect of intrafollicular injection of fulvestrant (an antagonist of ESRs) on follicular development and transcript abundance in granulosa cells. Methods Granulosa cells were obtained from the two largest follicles around follicular deviation, after FSH treatment and after intrafollicular injection of fulvestrant. Ovarian follicular dynamics monitored by ultrasonography and quantitative real time PCR were used to validate the in vivo model and investigate the effects of FSH supplementation or ESR blockade on mRNA expression of estradiol-related genes. Results ESR1 and ESR2 were expressed in granulosa cells of both dominant (F1) and subordinate (F2) follicles, but their transcripts levels were higher in F1 than F2 after follicular deviation. FSH treatment maintained mRNA levels of both ESR1 and ESR2 in F2 follicles at similar levels observed in F1 follicles. Intrafollicular injection of 100 μM fulvestrant inhibited follicular growth and decreased CYP19A1 mRNA levels. Transcript levels for both ESR1 and ESR2 were not affected by fulvestrant injection. Analyses of FSH-regulated genes revealed that ESRs inhibition in the dominant follicle decreased the transcript levels of the GJA1 but not those of PRKAR2B, MRO or LRP11 genes. Conclusions Our findings indicate that: both ESR1 and ESR2 are regulated during follicular deviation and dominance in cattle and in response to FSH treatment, and ESRs are required for normal gene expression and development of the dominant follicle. Furthermore, we have validated an in vivo model to study estrogen signaling during follicular development that allows paracrine signaling between different follicular cells in a physiological endocrine environment. [ABSTRACT FROM AUTHOR]

The objective of this study was to characterize the profiles of Ang-(1-7), MAS receptor, ACE2, NEP and PEP during the ovulatory process in cattle. For this study, 40 synchronized cows with follicular diameter ≥ 12 mm were ovariectomized at different time-points (0, 3, 6, 12 and 24 h) after i.m. application of gonadotropin-releasing hormone (GnRH) to induce a luteinizing hormone surge. Follicular fluid was collected for measuring Ang-(1-7) by radioimmunoassay. Theca and granulosa cells were isolated from the preovulatory follicles to evaluate the gene expression of MAS receptor, ACE2, NEP and PEP by qRT-PCR assay. Cross-contamination between theca and granulosa cells was tested by RT-PCR to detect cytochrome P450 aromatase (CYP19A1) and 17α-hydroxylase (CYP17A1) mRNA. Ang-(1-7) levels were constant until 12 h and then increased (p < 0.05) at 24 h after GnRH. Messenger RNA expression of MAS, ACE2, NEP and PEP was detected in theca and granulosa cells at all time-points after GnRH. In granulosa cells, ACE2, NEP and PEP were differentially expressed after GnRH treatment (p < 0.05). In conclusion, the Ang-(1-7), MAS receptor, ACE2, NEP and PEP profiles in preovulatory follicles indicate that Ang-(1-7) plays a role in the regulation of the ovulatory process in cattle. [ABSTRACT FROM PUBLISHER]

This aim of this study was to investigate the effectiveness of intravaginal devices (IVDs), containing medroxyprogesterone acetate (MPA), for controlling the estrous cycle in pubertal gilts. Gilts were assigned to four treatments: Control (no IVD); IVD containing 100 (IVD100), 200 (IVD200) or 400 mg (IVD400) of MPA. The IVDs were inserted on day 12 of the estrous cycle and maintained intra-vaginally for 14 days. The percentage of gilts in estrus, interval between IVD removal and estrous onset, adjusted farrowing rate (AFR), total number of piglets born (TPB), follicle size and serum progesterone (P4) were recorded. None of the gilts expressed estrus during the IVD treatment period. All gilts of the control group expressed estrus (15/15; 100%) which was greater (P = 0.03) than all IVD-treated gilts (33/44; 75%); however, there were no treatment differences (P = 0.09). The interval between IVD removal and estrous onset was shorter for IVD100 (3.8 ± 0.6 d) compared to IVD400 (5.3 ± 0.6 d; P = 0.05). The IVD400-treated gilts had smaller follicles than the IVD100-treated gilts (P = 0.05). The P4 concentrations were similar among treated groups (P = 0.99). The AFR did not differ among treatment groups (P = 0.37); however, the control group had a greater TPB than the other treatment groups (P = 0.04). The gilts treated with IVDs had longer interval to estrous expression. The most effective dosage was 400 mg of MPA, considering both the minimal follicular growth during the IVD treatment period and the lesser numbers of persistent follicles. [ABSTRACT FROM AUTHOR]

Treating lactating sows with chorionic gonadotropins may allow controlling their post-weaning reproductive function, despite the occurrence of anestrous during lactation. This article reviews the potential effectiveness of treatment with both equine and human chorionic gonadotropins (eCG and hCG, respectively) during lactation on the control of estrus expression and ovulation in weaned sows. The use of 1,000 IU hCG at 24 and 48 h postpartum may induce ovulation in the treated sows, but the ovulation rate may be variable. Pregnancy rates may be improved with combined treatment after the second week of lactation with both chorionic gonadotropins: 1,500 IU eCG plus 500 - 1,000 hCG; or 1,000 IU eCG plus 1,000 IU hCG. Treatment with eCG (1,000 - 2,000 IU) at the end of lactation may result in acceptable estrus expression and ovulation rates, although with marginal benefit for pregnancy rates. The subsequent response to treatments with chorionic gonadotropins during lactation is likely influenced by the treatment period, the suckling frequency during lactation, and the boar exposure during the weaning-to-estrus interval. A better understanding of the efficiency of such steroid-free treatments is increasingly relevant due to the constraints of the use of steroid hormones in livestock reproductive management., Competing Interests: Conflicts of interest: The authors have no conflict of interest to declare., (Copyright © The Author(s).)

The impact of GnRH treatment on the day of TAI in beef cows has received limited investigation, especially concerning its association with estrus expression. Consequently, two experiments were conducted to assess the potential of GnRH treatment on the day of TAI to enhance fertility according to the expression or not of estrus in beef cows. Experiment 1 aimed to determine ovulation rate and luteal function, while Experiment 2 aimed to determine the effect of the two GnRH treatment approaches on pregnancy rate. In Experiment 1, multiparous Brangus suckling cows (n = 17) were submitted to an 8-day TAI protocol. Estrus occurrence was evaluated based on chalk removal on D10 (TAI) and cows were assigned to receive GnRH (25µg lecirelin; im) according to the group: GnRH (n = 7), regardless of estrus expression; or selectGnRH (n = 10), only cows not detected in estrus. Ovulation rate occurring until 77h after IVD removal did not differ (p = 0.17) between GnRH (85.7%; 6/7) and selectGnRH (100%; 10/10). Also, corpus luteum size and serum progesterone concentration were not affected (p>0.05) by treatments. In Experiment 2, crossbred taurine suckled cows (n = 384) were submitted to the same protocol as described in Experiment 1 and were randomly allocated to GnRH or selectGnRH groups. There was no difference in P/AI between groups (selectGnRH = 55.6%; GnRH = 54.3%; p = 0.7) 30 days after TAI. As expected, there was a pronounced effect (p<0.0001) of estrus expression on P/AI (Estrus = 61.5%; No estrus = 33.0%), regardless of group. In summary, ovulation timing and rate and luteal function did not differ between groups. Also, GnRH administration only in cows that do not show estrus is recommended, considering hormone savings and similar conception rate., Competing Interests: Conflicts of interest: The authors have no conflict of interest to declare.

The aim of this study was to investigate the association between chronic Anaplasma marginale and Babesia spp. infection and hematological parameters of pregnant and non-pregnant taurine heifers. Blood samples from 94 females were collected on the first day (D-10) of timed artificial insemination (TAI) protocol and on pregnancy diagnosis (D+34). Hematological parameters were determined and compared between pregnant (PG) and non-pregnant (NPG) heifers, and within group at different sampling days. Real-time PCR (qPCR) was used to determine A. marginale and Babesia bovis infection, and for absolute quantification of Babesia spp. between PG and NPG groups. Correlation analysis was performed between the number of gDNA copies (CN) of Babesia spp. and hematological parameters. On D-10, mean hemoglobin concentration was higher for NPG, and hematocrit and total plasma protein were higher on D+34 for both groups. There was no difference in Babesia spp. CN between groups. In the first qPCR, all heifers were positive for A. marginale and B. bovis. Significant correlations were found between hemoglobin and erythrocyte and between hemoglobin and hematocrit (r = 0.8082 and r = 0.3009, respectively). Low levels of A. marginale and Babesia spp. did not affect hematological parameters of chronically infected pregnant and non-pregnant taurine heifers.

Radiotherapy causes destruction of tumor cells, but also threatens the integrity and survival of surrounding normal cells. Then, woman submitted to irradiation for cancer treatment may present permanent ovary damage, resulting in impaired fertility. The objective of this study was to investigate the effects of therapeutic doses of ionizing radiation (IR), used for ovarian cancer treatment in humans, on bovine cumulus-oocyte complexes (COCs) as experimental model. Bovine ovaries were exposed to 0.9 Gy, 1.8 Gy, 3.6 Gy or 18.6 Gy IR, and then COCs were collected and used to evaluate: (a) oocyte nuclear maturation; (b) presence of phosphorylated H2A.X (γH2AX), as an indicator of DNA double-strand breaks (DSBs); and (c) expression of genes involved in DNA repair ( TP53BP1, RAD52, ATM, XRCC6 and XRCC5 ) and apoptosis ( BAX ). The radiation doses tested in this study had no detrimental effects on nuclear maturation and did not increase γH2AX in the oocytes. However, IR treatment altered the mRNA abundance of RAD52 (RAD52 homolog, DNA repair protein) and BAX (BCL2-associated X protein). We conclude that although IR doses had no apparent effect on oocyte nuclear maturation and DNA damage, molecular pathways involved in DNA repair and apoptosis were affected by IR exposure in cumulus cells., Competing Interests: Conflicts of interest: The authors have no conflict of interest to declare.

The identification of mutations in the genes encoding bone morphogenetic protein 15 (BMP15) and growth and differentiation factor 9 (GDF9) associated with phenotypes of sterility or increased ovulation rate in sheep aroused interest in the study of the role of local factors in preantral and antral folliculogenesis in different species. An additive mutation in the BMP15 receptor, BMPR1b, which determines an increase in the ovulatory rate, has been introduced in several sheep breeds to increase the number of lambs born. Although these mutations indicate extremely relevant functions of these factors, the literature data on the regulation of the expression and function of these proteins and their receptors are very controversial, possibly due to differences in experimental models. The present review discusses the published data and preliminary results obtained by our group on the participation of local factors in the selection of the dominant follicle, ovulation, and follicular atresia in cattle, focusing on transforming growth factors beta and their receptors. The study of the expression pattern and the functionality of proteins produced by follicular cells and their receptors will allow increasing the knowledge about this local system, known to be involved in ovarian physiopathology and with the potential to promote contraception or increase the ovulation rate in mammals., Competing Interests: Conflicts of interest: The authors have no conflict of interest to declare.

Estradiol cypionate (EC) or GnRH have been widely used for ovulation induction in timed embryo transfer (TET). EC administration increases the proportion of cows that show estrus, whereas GnRH promotes more synchronized ovulations. The aim of the present study was to evaluate the potential beneficial effects of combining EC and GnRH in TET. In experiment 1, no difference was observed on serum progesterone concentrations on Day 6 and 13 after GnRH treatment between GnRH and EC+GnRH groups. In experiment 2, pregnancy per embryo transfer (P/ET) did not differ ( p = 0.69) between GnRH (62.8%) and EC+GnRH (58.7%) groups. In conclusion, combining EC and GnRH for ovulation induction does not increase progesterone secretion and pregnancy rate after TET in cattle., Competing Interests: Conflicts of interest: The authors have no conflict of interest to declare., (Copyright © The Author(s).)

Associations of the activity of the paraoxonase 1 (PON1) enzyme with boar sperm quality still needs to be characterized, since boar ejaculates present distinct portions with differences in sperm concentration and quality. This study evaluated PON1 activity in the serum, in the distinct portions of boar ejaculates and estimated correlations with sperm quality parameters. Ejaculates and blood samples were collected from six boars for three weeks (two per week per boar; n = 36). Serum and post-spermatic portion PON1 activities were positively correlated ( P = 0.01) but were both uncorrelated with the PON1 activity in the sperm-rich portion and in the whole ejaculate ( P > 0.05). Differences in PON1 activity among boars were only observed in the sperm-rich portion of the ejaculate ( P < 0.05). The PON1 activity in the serum and in the post-spermatic portion was generally negatively correlated with parameters of spermatozoa kinetics ( P < 0.05). In the sperm-rich portion, PON1 activity was positively correlated with sperm concentration ( P < 0.0001), curvilinear distance and velocity (both P < 0.05) and DNA integrity ( P < 0.05), but negatively correlated with straightness and linearity ( P < 0.05). Thus, boar ejaculates with increased PON1 activity in the sperm-rich portion may present increased concentration and spermatozoa with acceptable curvilinear velocity and distance and DNA integrity, which suggests that PON1 activity may be a biomarker for potential fertility., Competing Interests: Conflicts of interest: The authors have no conflict of interest to declare.

The aim of this study was to evaluate the long-term effects of recombinant bovine somatotropin (rbST) on follicle population and ovulatory follicle development in non-lactating dairy cows. Twenty-one Jersey cows were allocated in rbST (n=11) or control (n=10) groups. On day -60, cows in rbST group received 500 mg of somatotropin (s.c. Lactotropin®, Elanco). On day 0, control and rbST cows received an intravaginal progesterone-releasing device (1.9 g, CIDR ® , Zoetis) and GnRH (100 mg, IM, Factrel ® , Zoetis). On day 8, cows received PGF2α (25 mg, IM, Lutalyse®, Zoetis) and the CIDR ® was removed. Twelve hours after device removal (D8), serum, follicular fluid and granulosa cells samples were collected. Serum and follicular concentration of estradiol (E2) and progesterone (P4) were analyzed. Total RNA was extracted from granulosa cells to measure gene expression of LHCGR , STAR , HSD-3B1 , CYP11A1 , CYP19A1 , CYP17A1 , IGFR and PAPPA by real-time PCR. Ultrasonography was performed on days -60, -53, -46, -14, -7, 0 and 8 for antral follicle count. Results were analyzed by repeated measures ANOVA and t-test. There was no effect of rbST treatment on the number of follicles during the 60 days period, as well as no effect on serum and follicular fluid E2 and follicular fluid P4 at the moment of follicle aspiration. There was a reduction in PAPPA ( P = 0.006), CYP11A1 ( P = 0.04) and CYP19A1 ( P = 0.002) mRNA levels in granulosa cells of the pre-ovulatory follicle of rbST treated cows. In conclusion, a single dose of rbST did not have long-term effects on antral follicle population, serum and follicular E2/P4 concentrations in non-lactating dairy cows. Despite that, rbST injection decreased granulosa cell expression of genes related to steroidogenesis in the pre-ovulatory follicle., Competing Interests: Conflict of interest: The authors have no conflict of interest to declare., (Copyright © The Author(s).)

The transforming growth factors beta (TGFβ) are local factors produced by ovarian cells which, after binding to their receptors, regulate follicular deviation and ovulation. However, their regulation and function during corpus luteum (CL) regression has been poorly investigated. The present study evaluated the mRNA regulation of some TGFβ family ligands and their receptors in the bovine CL during induced luteolysis in vivo . On day 10 of the estrous cycle, cows received an injection of prostaglandin F2α (PGF) and luteal samples were obtained from separate groups of cows (n= 4-5 cows per time-point) at 0, 2, 12, 24 or 48 h after treatment. Since TGF beta family comprises more than 30 ligands, we focused in some candidates genes such as activin receptors ( ACVR-1A , -1B , -2A , -2B ) AMH , AMHR2 , BMPs ( BMP-1 , -2 , -3 , -4 , -6 and -7 ), BMP receptors ( BMPR-1A , -1B and - 2 ), inhibin subunits ( INH-A , -BA , -BB ) and betaglycan ( TGFBR3 ). The mRNA levels of BMP4 , BMP6 and INHBA were higher at 2 h after PGF administration (P<0.05) in comparison to 0 h. The relative mRNA abundance of BMP1 , BMP2 , BMP3 , BMP4 , BMP6 , ACVR1B , INHBA and INHBB was upregulated up to 12 h post PGF (P<0.05). On the other hand, TGFBR3 mRNA that codes for a reservoir of ligands that bind to TGF-beta receptors, was lower at 48 h. In conclusion, findings from this study demonstrated that genes encoding several TGFβ family members are expressed in a time-specific manner after PGF administration., Competing Interests: Conflicts of interest: The authors have no conflict of interest to declare., (Copyright © The Author(s).)