Plasma Estradiol Profile After Administration of Different Types of Estradiol Esters in Acyclic Mares.

Although the use of different estradiol esters has been extensively studied in hormonal protocols in cows, such information is lacking in mares. The present study aimed to assess the effects of treatment with the same doses and administration frequency of estradiol cypionate, estradiol benzoate and 17ß estradiol on plasma estradiol (E2) concentrations of acyclic mares and correlate the E2 profile to the endometrial edema score. Sixteen treatments were performed in 14 mares randomly divided into three groups: EB (n = 5), EC (n = 5), and 17ß (n = 6), receiving 10 mg on day 0 (D0), 6 mg on D1, and 4 mg on D2 of estradiol benzoate, estradiol cypionate, and estradiol 17ß, respectively. Blood samples, rectal palpations, and ultrasound evaluations were performed once daily, starting before the first estradiol treatment (D0) until edema disappearance or D8. Moderate to high edema was observed in all groups 24 hours after the first estradiol administration. Edema persisted above score 2 until D7, D5, and D4 in groups EC, EB, and 17ß, respectively. Higher edema was found on D2 in EB group compared to 17ß, on D6 and D7 in EC compared to 17ß, and on D8 in EC compared to EB (p ≤ .05). Maximum E2 concentrations were detected on D1 in groups: EB, showing a sharp decrease from D2 to D3 (p < .0001); and 17ß, where no differences were observed between treatment days (p ≥ .05). In the EC group, maximum concentrations were observed on D2, which remained high from D2 to D4 compared to the other days (p < .0001). Plasma concentration of E2 was higher in EB than the other groups on D0 and D1, and EB>EC>17ß on D2. Plasma E2 concentrations and edema score were positively correlated, being moderate in groups EB and EC and weak in the 17ß group. In conclusion, the most pronounced plasma E2 concentration was observed in EB group 24 hours after the administration of the first dose. Estradiol concentrations peaked 48 hours after EC administration, while a distinct peak was not observed in a 24h interval evaluation in the 17ß group. Moreover, high edema does not necessarily reflect on high E2 concentrations in acyclic mares.

Peri-estrus ovarian, uterine, and hormonal variables determine the uterine luminal fluid metabolome in beef heifers.

In cattle, uterine luminal fluid (ULF) is the main source of molecules that support embryo development and survival during the peri-implantation period. Our overarching hypothesis is that peri-estrus changes in uterine function, including ULF accumulation and absorption, are uneven among individuals, and affect ULF composition and fertility. Our objectives were (1) to characterize temporal and spatial changes in ULF volume, endometrial and luteal blood perfusion, endometrial and luteal size, and circulating progesterone concentrations during the peri-estrus period in beef heifers and (2) to associate such changes with the metabolite composition in the ULF, 4 days after estrus (d 0). Fourteen Bos indicus heifer that presented a PGF2α responsive CL received 500 µg PGF2α analog i.m. and were examined daily by rectal B-mode and pulse-wave color-Doppler ultrasonography until the fifth day after estrus (d 5). The composition of the ULF was analyzed by targeted mass spectrometry on d 4. Multivariate analyses clustered heifers according to ovarian, uterine, and hormonal variables in clusters A (n = 5) and B (n = 8 heifers). Concentrations of Pro, Ala, Leu, Gly, Val, Lys, Ile, Phe, Asp, Orn, Tyr, Arg, Trp, Suc, Cit, ADMA, the sum of essential Amino Acids (AA), sum of nonessential AA, sum of aromatic AA, and total AA were greater in cluster A (FDR ≤ 0.05). ULF volume dynamics and uterine, ovarian, and hormonal variables during the peri-estrus period presented a concerted variation among heifers within clusters, which was associated with the ULF composition 4 days after estrus.

Use of pregnancy-associated plasma protein-A during oocyte in vitro maturation increases IGF-1 and affects the transcriptional profile of cumulus cells and embryos from Nelore cows.

Insulin-like growth factor 1 (IGF-1) activity is established by the regulation of IGF binding protein activity, which blocks IGF-1 functions, whereas pregnancy-associated plasma protein-A (PAPP-A) improves IGF-1 bioavailability and facilitates binding to IGF receptors. To further extend our understanding of the effect of exogenous PAPP-A on bovine embryo production, we added this protein during in vitro maturation of cumulus-oocyte complexes (COCs); moreover, we assessed its effects on IGF-1 quantity in the maturation medium, embryonic yield and postwarming survival, blastocyst quality, and transcript abundance. Bovine COCs were matured in a serum-free medium, either with PAPP-A supplementation (100 ng/ml) or without (control). The treatment group produced higher IGF-1 concentrations in the maturation medium; however, showed no difference on cleavage, blastocysts rates, and embryonic survival 3 and 24 hr postcryopreservation. Regarding gene expression, VNN1 was upregulated, whereas AGPAT9, FASN, EGFR, HAS2, and IMPDH1 were downregulated in PAPP-A treated. PAPP-A treated, CPT2, DNMT3A, and TFAM were upregulated, whereas ATF4 and IFITM3 were downregulated. We concluded that although the addition of PAPP-A did not affect embryo yield and blastocyst survival, higher IGF-1 levels may affect embryo competence through differential expression of genes involved in lipid metabolism, oocyte competence, and mitochondrial function.

B-mode ultrasonography and gray scale histogram for evaluation of the nuchal ligament in Quarter horse / Ultrassonografia modo-B e histograma em escala de cinza para avaliação do ligamento nucal em equinos da raça Quarto de Milha

The gray scale histogram (GSH) makes it possible to measure the amount and distribution of gray shade frequencies, providing quantitative information on both echogenicity and echotexture of tissues. There is a need to diminish the subjectivity of the ultrasound images of the nuchal ligament (NL). This work proposes to evaluate the NL by ultrasound B-mode GSH images in different ages of Quarter horses. It used 15 healthy Quarter horses, which were classified by age into three different groups: "baby" (1), "sobreano" (2) and "adult" (3). The animals were submitted to chemical restraint for the ultrasound examination. Subsequently, the GSH tool was used in each image for statistical analysis. There was a significant difference between Mean and Mode between groups. Group 1 differed significantly when compared to Group 2 and 3. Group 2 presented superior echogenicity to Group 1 and 3. The height of the NL did not vary considerably between groups. GSH indicated that the echogenicity of NL in Quarter horses varies with age.(AU)

O histograma em escala de cinza (HEC) possibilita a mensuração da quantidade e distribuição da frequência de tonalidades de cinza, fornecendo informações quantitativas, tanto sobre a ecogenicidade quanto a ecotextura dos tecidos. Havendo a necessidade de diminuir a subjetividade das imagens ultrassonográficas do ligamento nucal (LN), esse trabalho propôs avaliar o LN por imagens ultrassonográficas modo-B pelo HEC em diferentes idades de cavalos da raça Quarto de Milha. Utilizou 15 cavalos da raça Quarto de Milha saudáveis os quais foram classificados por idade em três grupos diferentes: "baby" (1), "sobreano" (2) e "adulto" (3). Os animais foram submetidos a contenção química para a realização do exame ultrassonográfico. Posteriormente, a ferramenta HEC foi empregada em cada imagem para análise estatística. Houve diferença significativa entre as variáveis "Mean" e "Mode" entre os grupos. O Grupo 1 diferenciou significativamente quando comparado ao Grupo 2 e 3. O Grupo 2 apresentou ecogenicidade superior ao Grupo 1 e 3. Quanto à altura do LN não teve variação considerável entre os grupos. O HEC indicou que a ecogenicidade do LN em cavalos Quarto de Milha varia conforme a idade.(AU)

Supplementation with sunflower seed increases circulating cholesterol concentrations and potentially impacts on the pregnancy rates in Bos indicus beef cattle.

We aimed to evaluate the effect of supplementation with sunflower seed on blood concentrations of progesterone and cholesterol and on the pregnancy rate in beef cattle subjected to timed artificial insemination (TAI) and timed embryo transfer (TET). In experiment 1, cows were received 22-day supplements containing (sunflower, n = 66) sunflower seed or not (control, n = 67) immediately after a progesterone/estradiol-based TAI protocol (Day 0). The cholesterol concentration on Day 21 and the pregnancy rate were greater (P < 0.03) in the sunflower group (148.2 ± 6.1 mg/dL and 66.7%) than those in the control group (116.0 ± 6.4 mg/dL and 47.8%). In experiment 2, heifers received an in vitro-produced embryo 7 days after the expected time of the synchronized ovulation. Heifers were separated into two supplementation groups (sunflower, n = 106 and control, n = 111) for 22 days. The plasma progesterone concentration on Day 7 was not different between the groups. However, on Day 19, the plasma progesterone concentration was greater (P < 0.0001) in the sunflower group (5.8 ± 0.4 ng/mL) than that in the control group (3.5 ± 0.4 ng/mL). A greater (P < 0.05) cholesterol concentration was observed in the sunflower group than that in the control group on Days 7 (306.0 ± 11.6 vs. 277.1 ± 11.9 mg/dL, respectively) and 19 (260.5 ± 8.0 vs. 232.0 ± 8.0 mg/dL, respectively). The pregnancy rate was greater (P = 0.01) in the sunflower-treated heifers (55.7%) than that in control-treated heifers (36.9%). Results indicate that sunflower seed supplementation increases the circulating cholesterol concentrations and potentially impacts the pregnancy rate in suckled beef cattle subjected to TAI or TET.

Plasma steroid dynamics in late- and near-term naturally and artificially conceived bovine pregnancies as elucidated by multihormone high-resolution LC-MS/MS.

The plasma levels of corticosteroids and sex steroids during pregnancy are key indicators of mammalian placental function and the onset of parturition. Steroid hormones are believed to be disturbed in pregnancies produced using assisted reproductive technologies (ARTs) due to placental dysfunction and the frequently observed lack of parturition signals. To elucidate the plasma steroid dynamics, a liquid chromatography-tandem mass spectrometry method was developed and used to determine the levels of corticosteroids (corticosterone, 11-deoxycortisol, and cortisol) and their direct precursors (progesterone and 17α-OH-progesterone) as well as sex steroids (androstenedione, estrone, estrone sulfate, testosterone, and 17ß-estradiol) in bovine plasma. The levels of these 10 steroids in recipient cows carrying naturally conceived (control), in vitro fertilized (IVF), or cloned (somatic cell nuclear transfer) conceptuses were compared during late-term pregnancy (30 days before parturition), during near-term pregnancy (1 day before parturition), and on the day of parturition (day 0). Significant differences were observed among the corticosteroid levels: higher levels of corticosterone, 11-deoxycortisol, and cortisol were detected in cloned pregnancies at day 30; lower levels of corticosterone were observed in ART-derived pregnancies at days 1 and 0; and estrone and estradiol levels were higher in IVF pregnancies throughout the final development. These results suggested an upregulation of the P450C11 and P450C21 enzymes 30 days before parturition in somatic cell nuclear transfer pregnancies and an overactivation of the aromatase enzyme in IVF pregnancies. Taken together, the monitoring of multiple steroid hormones revealed that the pregnancies obtained using ART exhibited plasma steroid concentration dynamics compatible with the dysregulation of steroidogenic tissues.

Changes in luteinizing hormone secretion after estradiol treatment in prepubertal Nelore heifers.

Changes in luteinizing hormone (LH) secretion after 17ß-estradiol (E(2)) injection were evaluated during sexual maturation in 10 prepubertal Nelore heifers. Heifers were divided into 2 groups: intact (I) and ovariectomized (OVX). 17ß-estradiol (2 µg/kg) was administered to both groups at 10, 13, and 17 mo of age. Only at 10 mo of age was there a greater mean LH concentration in OVX heifers (1.33 ± 0.29 ng/mL) compared with the I group (0.57 ± 0.15 ng/mL). At 13 and 17 mo of age there was no significant difference between the 2 groups in any of the evaluated variables (number of peaks, total peak area, greatest peak area, and time to greatest peak occurrence). This suggests a decrease in negative E(2) feedback associated with an increase in positive feedback to LH secretion during sexual maturation, and these were likely the key factors that determined the time of first ovulation in Nelore heifers.

Ovariectomy by left flank approach in prepubertal Nelore (Bos indicus) heifers.

The main objective of this study was to evaluate a recumbent left flank approach to bilateral ovariectomy in prepubertal heifers and to develop an optimal surgical technique for this procedure. Both ovaries were removed from 6 Nelore heifers by left flank approach without any complications, except in 1 heifer, which was believed to have had only 1 ovary based on ultrasound and exploration during surgery, but was later found to have a remaining functional ovary. Ovariectomy via left flank approach in recumbent prepubertal heifers is feasible and technically easy. This procedure does not involve special instrumentation and, despite the invasive approach, it allows optimal visualization of the ovaries and uterus.

Follicular dynamics and plasma FSH and progesterone concentrations during follicular deviation in the first post-ovulatory wave in Nelore (Bos indicus) heifers.

The objective of the present study was to characterize ovarian follicular dynamics and hormone concentrations during follicular deviation in the first wave after ovulation in Nelore (Bos indicus) heifers. Ultrasonographic exams were performed and blood samples were collected every 12h from the day of estrus until 120-144 h after ovulation in seven females. Deviation was defined as the point at which the growth rate of the dominant follicle became greater than the growth rate of the largest subordinate follicle. Deviation occurred approximately 65 h after ovulation. Growth rate of the dominant follicle increased (P<0.05) after deviation, while growth rate of the subordinate follicle decreased (P<0.05). Diameter of the dominant follicle did not differ from the subordinate follicle at deviation (approximately 5.4mm). The dominant follicle (7.6mm) was larger (P<0.05) than the subordinate follicle (5.3mm) 96 h after ovulation or 24h after deviation. Plasma FSH concentrations did not change significantly during the post-ovulatory period. The first significant increase in mean plasma progesterone concentration occurred on the day of follicular deviation. In conclusion, the interval from ovulation to follicular deviation (2.7 days) was similar to that previously reported in B. taurus females, but follicles were smaller. Diameters of the dominant follicle and subordinate follicle did not differ before deviation and deviation was characterized by an increase in dominant follicle and decrease in subordinate follicle growth rate. Variations in FSH concentrations within 12-h intervals were not involved in follicular deviation in Nelore heifers.