Follicular Dynamics Research Articles

Open Cows in a Beef Cattle Enterprise Managed in a Seasonal Breeding Program? An Appraisal of Their Reproductive Performance Based on Body Fat Reserves

A comparison between two distinct levels of back fat thickness in open cows, females not becoming pregnant during the previous breeding season, was carried out in order to evaluate their reproductive performance prior to the next natural mating in a seasonal breeding program. Thirty open cows from the previous breeding season were divided evenly by age and back fat thickness (BFT) in two groups. HBFT (high back fat thickness ≥ 70 mm) and LBFT (low back fat thickness ≤ 70 mm), values statistically different between groups (P < 0.05). The study was divided into three phases in which the formation of CL and follicular dynamics were recorded by ultrasound and progesterone. Also, serial monitoring of glucose, urea and triglycerides and back fat thickness were studied. A fertile bull previously approved from a breeding soundness evaluation was introduced after the third phase and remained with the herd for 90 days. No differences were found in the follicular dynamics; the number of animals cycling in the two groups and fertility were also similar (80%). Differences in urea and triglycerides were found in cows with high scores of BFT. In conclusion, the decision of keeping open cows to the next breeding season must be based on an economical advantage to the farmer.

Glycerin supplementation strategies for three or seven days affects oxidative stress, follicle dynamics and ovulatory response in Morada Nova sheep.

This study examined the effect of glycerin supply strategies in different short-term protocols on follicular dynamics and ovulatory rate in Morada Nova sheep. Eighteen Morada Nova ewes with body condition > 2.9 had their estrus and follicular waves synchronized using three injections of prostaglandin analogue at seven-day intervals. All animals received the same diet during 21 days, which consisted of a total mixed ration (TMR) based on chopped elephant grass and concentrate twice daily. In the control group (n=9), ewes were fed the TMR diet. In the other four groups, ewes received 150 mL of glycerol daily, supplied as an oral drench or mixed in the TMR during three or seven days prior to the application of the third PGF2 alfa analogue. These groups were named as follows: Drench3d (n=10), Drench7d (n=8), TMR3d (n=9) and TMR7d (n=9). Follicle dynamics were monitored by ultrasonography, and plasma glucose and glutathione peroxidase levels were measured at the third prostaglandin administration. Six days after the final PGF2 alfa analogue dose, ovulatory rate was measured by laparoscopy. Glucose was higher (P< 0.001) in the glycerin-treated groups than in control group (83.7 ± 1.7 vs. 68.4 ± 4.5 mg. dL-1; P < 0.001). Ewes in the TMR3d, Drench7d and TMR7d groups had a greater (P < 0.001) number of large follicles (≥ 3 < 5 mm), and the presence of follicles larger than 5 mm was observed. In the same groups, at the third PGF2 alfa analogue dose, a greater (P < 0.001) number of growing follicles (> 3 mm) and a larger size of the largest follicle (P < 0.001) were also recorded. Ovulation rate was 30% higher in the groups that received glycerin for seven days (1.6 ± 0.1 53 vs. 1.1 ± 0.1; P < 0.05), and they also exhibited a 38% reduction in glutathione peroxidase. Thus, the use of glycerin in Morada Nova sheep as a source of energy in short-term supplementation for increase ovulation rate is an efficient strategy when provided for seven days, either orally or in the feed.

Fertility Response Following Estradoublesynch and Progesterone Based Ovsynch Protocols in Delayed Pubertal Buffalo Heifers

The study evaluated the fertility response following Estradoublesynch and progesterone-based Ovsynch protocols in delayed pubertal buffalo heifers. A total of twenty delayed pubertal buffalo heifers were subdivided in two equal subgroups. Group I animals were treated with PGF2α analogue (500 µg cloprostenol sodium) i.m. on day 0, GnRH analogue (20 µg buserelin acetate) on day 2, PGF2α analogue (500 µg cloprostenol sodium) repeated on day 9 and estradiol benzoate 1 mg intramuscular (i.m.) on day 10. Fixed-time artificial insemination (FTAI) was done at 48 and 60h. Group II animals were subjected to GnRH analogue (20 µg buserelin acetate) i.m. along with CIDR insert intravaginally on day 0, PGF2α analogue (500 µg cloprostenol sodium) i.m. and removal of CIDR on day 7, GnRH analogue on day 9 followed by FTAI at 12 and 24h. Blood collection (plasma P4) and ultrasonography (luteal and follicular dynamics) was performed on day 0, 9, 10, 12 in group I and day 0, 7, 9 and 10 in group II animals. Pregnancy diagnosis was made at 45-60 days post AI. Estrus response was observed in all the buffalo heifers of group I and II. First service conception rate was significantly higher in group I (50%) compared to group II (20%) (P&lt;0.05). Overall pregnancy rate was almost similar in both group I and II (60% vs 70%). In conclusion, estradoublesynch and progesterone-based ovsynch protocol can effectively induce ovulatory estrus and improve pregnancy rates in delayed pubertal buffalo heifers.

Ovarian follicular dynamics in peripubertal Malabari goats

The aim of this study was to characterise the follicular dynamics in first two oestrous cycles of peripubertal Malabari goats. Transrectal ultrasonography was carried out on alternate days of oestrous cycles after first observed oestrus. Follicular studies revealed presence of two to five wave cycles with predominant three and four wave patterns. The interwave interval for ovulatory wave was longer than preceding waves in three and five wave cycles. The diameter of largest follicles in wave 1 was larger than follicles of subsequent waves of three, four and five wave cycles. The largest follicles in each wave could be observed from day 3 to day 13 of oestrous cycle. Majority of ovulations occurred on day 20 of oestrous cycles and short cycles were more in first oestrous cycle. The overall mean per cent of single and double ovulations were 56.3 and 43.8 and mean ovulatory follicle size was 6.59 ± 0.11 mm in peripubertal goats. The mean length of first and second oestrous cycle was observed as 17.72± 0.79 and 20.19± 0.43 days, respectively and mean duration of first and second oestrus was 36.88 ± 2.44 and 40.13 ± 1.84 h, respectively. The study revealed that predominant wave pattern was three and four waves in peripubertal Malabari breeds. The first oestrous cycle length was shorter and characterised by higher percentage of single ovulations with smaller periovulatory follicle size. Larger ovulatory follicles and double ovulations were observed in second oestrous cycle.

Influence of eCG and reproductive management in the resynchronization of ovulation in dairy goats.

Resynchronization protocols have been proposed as a way of shortening females’ unproductive time in the flock, with good results in cattle and sheep. In goats, initial studies have shown that a second progestogen device inserted before luteolysis and pregnancy diagnosis does not interfere with the corpus luteum lifespan or functionality. This study aimed to evaluate the follicular growth, ovulation pattern and pregnancy rate after insertion of a second and new progestogen device for resynchronizing, with or without equine Chorionic Gonadotrophin (eCG), submitted to natural mating (NM) or artificial insemination (AI) to propose a viable resynchronization protocol for dairy goats. A total of 38 multiparous Saanen goats underwent a short-term progesterone protocol [six days exposed to medroxyprogesterone acetate (MAP) intravaginal sponges + 200 IU eCG and 0.12 mg of cloprostenol sodium on the 5th day + 0.025 mg of lecirelin 34 hours after sponge withdrawal] and, on day 16th after the ovulation, received a new MAP device which was retained until day 21. At this moment females were split into four groups: GeCG+NM – 100 IU eCG with NM; GSal+NM – saline solution with NM; GeCG+AI – 100 IU eCG with AI; and GSal+AI – saline solution with AI. Ultrasound scans were performed every 12 h from sponge withdrawal (day 21) until 108 h after sponge withdrawal (day 25) for follicular dynamics evaluation, at 240 h (day 31) for assessing the presence of active corpus luteum, and on day 60 for pregnancy diagnosis. No differences were found regarding ovulation time, synchronization and follicle size. However, GeCG+NM presented a greater estrus manifestation rate (100%) and pregnancy rate (62.5%) when compared to GSal+AI. In conclusion, resynchronization protocols in dairy goats may present satisfactory results.

Effect of ovarian follicular wave pattern and endocrine characteristics on pregnancy outcome in cows.

The effect of two (2W) versus three (3W) wave patterns of follicular dynamics and concurrent endocrine milieu of follicle-stimulating hormone (FSH), luteinizing hormone (LH), oestradiol 17-β (E2) and progesterone (P4) were investigated during one interoestrous interval (IEI) before insemination, on ensuing pregnancy, in 70 lactating Jersey crossbred cows. The findings were evaluated for between [included all (overall) 2W-O and 3W-O cows] and within [after separating pregnant (P) and non-pregnant (NP) cows in 2W and 3W] wave patterns. The propensity of two (58.6%, n=41) and three (41.4%, n=29) wave patterns was similar (p=.15). The IEI, shorter by 2.6days for 2W-O versus 3W-O (p<.0009), predicted wave pattern as 100% 2W-O cows had IEI of ≤21days, present only in 27.6% 3W-O cows (p<.0001). The ovulatory follicle persisted for a significantly shorter duration for 3W-O versus 2W-O cows. The average FSH, LH, E2 and P4 concentrations during the IEI did not differ for between and within the wave patterns. Pregnancy rate (%) of 58.6 versus 41.4 (p=.15) for 2W-O versus 3W-O and 56.1-P versus 43.9-NP (p=.44) for within 2W was similar, but tended to differ for within the 3W pattern (69.0-P versus 31.0-NP, p=.06). The pregnancy outcome was influenced by the age of ovulatory follicle for between the wave patterns and by follicular count as well as FSH surge concentration for within the wave patterns. A shorter luteal phase reduced the pregnancy outcome, a novel finding of the present study.

SOD1 Gene Silencing Promotes Apoptosis and Suppresses Proliferation of Heat-Stressed Bovine Granulosa Cells via Induction of Oxidative Stress.

Heat stress (HS) compromises dairy cattle reproduction by altering the follicular dynamics, oocyte maturation, and normal physiological function of ovarian granulosa cells (GCs), eventually resulting in oxidative damage and cell apoptosis. To protect the cells from oxidative damage, the Superoxide dismutase-1 (SOD1) degraded the hydrogen peroxide (H2O2) to oxygen (O2) and water. The objective of the current study was to investigate the impact of SOD1 silencing on intracellular ROS accumulation, cell viability, MMP, hormone synthesis (P4, E2), cell proliferation, and apoptosis in GCs under HS. The mechanistic role of SOD1 regulation in the heat-stressed GCs was explored. SOD1 gene was successfully silenced in GCs and confirmed at both transcriptional and translational levels. We found that silencing of SOD1 using siRNA under HS aggravated intracellular accumulation of reactive oxygen species, apoptosis, disrupted the mitochondrial membrane potential (MMP), altered transition of the cell cycle, and impaired synthesis of progesterone (P4) and estrogen (E2) in GCs. The associative apoptotic, steroidogenic, and cell cycle genes (BAX, Caspase-3, STAR, Cyp11A1, HSP70, PCNA, and CyclinB1) were used to confirm the results. These results identify a novel role of SOD1 in the modulation of bovine ovarian GC apoptosis, which provides a target for improving the fertility of heat-stressed dairy cows in summer.

Evaluation of ImageJ software in ultrasonic image analysis: Follicular and luteal morphological characteristics of cattle

This study was conducted to compare the effectiveness of two methodologies in evaluating B- and Doppler-mode ultrasonic images: analysis using ultrasonic software and utilizing a computer with ImageJ software. To determine if ImageJ software utilization is an efficacious alternative to ultrasonic software device- analysis, there were comparisons of values when using the two methods for morphological and vascular characteristics of follicular dynamics and luteal function in 18 crossbred cattle. From day 8 of an ovarian dynamics synchronization treatment regimen period until the time of ovulation (Day 10), B-mode and power-flow ultrasonic cineloops were obtained every 12 h to assess follicular diameter, wall area, and wall blood perfusion area. On Day 14 after ovulation, US cineloops of ovaries were obtained in B mode and power flow to evaluate various morphological and vascular characteristics of the corpus luteum (CL), including luteal diameter, luteal area, and CL blood perfusion area. Cineloops were evaluated and analyzed using ultrasonic software, and in a computer with ImageJ software. To evaluate consistency in results between the two methods, there was evaluation utilizing paired t-test, Pearson correlation coefficient, Bland-Altman plot, and Linear Regression Test procedures to calculate proportion of bias between values for measurements of variables evaluated. Results indicated none of the values for variables before and after ovulation differed (P > 0.05). It, therefore, was concluded that utilization of ImageJ software is an efficacious biomedical technique to analyze ultrasonic images of morphological and vascular characteristics before and after ovulation in cattle.

Follicular dynamics, luteal characteristics, and progesterone concentrations in synchronized lactating Holstein cows with high and low antral follicle counts

We performed two experiments in high milk-producing Holstein cows. First, we evaluated the repeatability of the antral follicle count (AFC) – where all follicles were ≥2 mm in diameter – during the days of the ovulation synchronization protocol for timed artificial insemination (TAI) in dairy cows (n = 20). In the second study we investigated the follicular dynamics and Doppler ultrasonography in cows with low (≤15 follicles, n = 9) and high (≥25 follicles, n = 9) AFC, and measured progesterone (P4) concentrations during CL evaluation. In addition, the diameter of the preovulatory follicle, area of blood flow of the preovulatory follicle, CL area, and CL blood flow were compared between groups with high and low AFC. Ultrasound examinations were performed during the days of the protocol and 10 days after ovulation to assess the CL. The constancy of the AFC was analyzed by calculating the repeatability. Parametric variables were analyzed using the t-test, and nonparametric variables were analyzed using the Mann-Whitney test (P ≤ 0.05). The repeatability of the AFC on different days of the protocol was 0.98. Cows with low AFC showed a greater diameter of the preovulatory follicle (16 ± 0.3 vs. 15 ± 0.4 mm), blood flow area of the preovulatory follicle (17.3 ± 2.3 vs. 8.1 ± 3.2 mm2), CL area (579.1 ± 16.9 vs. 405.8 ± 21.2 mm2), area of CL perfusion (97.1 ± 9.4 vs. 68.3 ± 5.2 mm2), as well as higher P4 levels (3.1 ± 0.2 vs. 2.2 ± 0.2 ng/mL). In high milk-producing Holstein cows, the AFC has high repeatability on different days of the TAI protocol, and cows with a low AFC show larger preovulatory follicles and CLs with greater perfusion, in addition to greater P4 concentrations in comparison to those cows with a high AFC.

Cathepsin B Regulates Mice Granulosa Cells' Apoptosis and Proliferation In Vitro.

Cathepsin B (CTSB), a lysosomal cysteine protease’s high expression and activity, has been reported to cause poor-quality embryos in porcine and bovine. Nevertheless, CTSB functions in mice granulosa cells remain to explore. To discuss the CTSB functional role in follicular dynamics, we studied apoptosis, proliferation, cell cycle progression, and related signaling pathways in primary mouse granulosa cells transfected with small interference RNA specific to CTSB (siCTSB) for 48 h. Further, mRNA and protein expression of cell proliferation regulators (Myc and cyclin D2), apoptosis regulators (caspase 3, caspase 8, TNF-α, and Bcl2), steroidogenesis-related genes (FSHR and CYP11A1), and autophagy markers (LC3-I and ATG5) were investigated. In addition, the effect of CTSB on steroidogenesis and autophagy was also examined. Flow cytometry analysis assay displayed that silencing of CTSB decreased the early and total apoptosis rate by downregulating TNF-α, caspase 8, and caspase 3, and upregulating Bcl2. By regulating Myc and cyclin D2 expression and activating the p-Akt and p-ERK pathways, CTSB knockdown increased GC proliferation and number. A significant decline in estradiol and progesterone concentrations was observed parallel to a significant decrease in autophagy-related markers LC3-I and ATG5 compared to the control group. Herein, we demonstrated that CTSB serves as a proapoptotic agent and plays a critical role in folliculogenesis in female mice by mediating apoptosis, autophagy, proliferation, and steroidogenesis. Hence, CTSB could be a potential prognostic agent for female infertility.

The antioxidant effect of Michauxia campanuloides on rat ovaries

This study investigated how Michauxia campanuloides affects the Proliferating Cell Nuclear Antigen (PCNA) expression in granulosa cells, the ovarian histomorphology and serum total antioxidant capacity (TAC) in rats. Rats were divided into control (C), treatment 1 (T 1) and treatment 2 (T 2) groups. The rats in the T 1 and T 2 groups received aqueous extract of M. campanuloides at doses of 20 mg/kg/day and 40 mg/kg/day orally for 21 days, respectively. Serum TAC levels, follicles counts including primordial, primary, preantral, antral and atretic follicles, and PCNA expression in granulosa cells were evaluated. Numbers of preantral follicles increased in T 1 and T 2 groups compared to C group (P &lt; 0.05). TAC and numbers of preantral and antral follicles increased in T 2 group compared to T 1 and C groups (P &lt; 0.05). PCNA expression in granulosa cells was increased in T 2 group compared to T 1 and C groups (P &lt; 0.01). In conclusion, treatment with M. campanuloides had positive effects on antioxidant activity, follicular dynamics, and PCNA expression of granulosa cell in rats.

Periodic critical micro-nutrients supplementation affects reproduction performance in peri-parturient dairy buffaloes

The study aimed to evaluate the effect of critical micronutrient (Zn, Cu, Co, Cr, Se, vitamin E, vitamin A, and niacin) supplementation at the weekly interval during the periparturient period (56 days prepartum till 56 days post-partum) on reproduction status of Murrah buffaloes. The experiment was performed on 22 healthy Murrah buffaloes after dividing them into two equal groups, representing control and treatment, respectively. A feeding schedule of both the groups was the same except that additional micronutrient supplementation was carried out once in a week in buffaloes of treatment group after proper mixing with concentrate mixture. Ultrasonography was performed once in a month for initial 90 days postpartum for assessment of cyclicity status of buffaloes. Treatment was unable to alter the cyclicity status of buffaloes for the initial 90 days but improved the numbers of large and medium-size follicles during the due course of the study. It may be concluded that weekly supplementation of critical micronutrients during the periparturient period is helpful in improving the follicular dynamics.

Exogenous progesterone-dependent modulation in the follicular dynamics of Bos indicus cattle undergoing repeated ovum pick-up sessions.

The objective of the present study was to evaluate the role of serum progesterone (P4) in follicular dynamics, oocytes' recovery and quality and their in vitro developmental competence during consecutive ovum pick-up (OPU) sessions in Bos indicus dairy cows. Wave-synchronized Sahiwal cattle (n=20) were randomly divided into treatment (n=10) and control (n=10) groups. CIDR was used as a source of external progesterone in the treatment group. Four consecutive OPU sessions at 96-hr intervals were conducted and repeated ultrasonography at 12-hr intervals was done to monitor follicular dynamics. The viable oocytes were processed for IVC following IVM and IVF until day 7. The serum P4 concentrations in the P4 and control groups were recorded as 2.31±0.059 versus.0.32±0.065ng/ml, respectively (p<.05). In the treatment group, the total number of recorded follicles was higher (p<.05; 12.05±0.37 versus. 10.87±0.40), whilst the growth rate (mm/day) of follicles was lower (p>.05). Per session recovered oocytes (5.31±0.19 versus. 3.58±0.21; p<.0001) and recovery rate (54.23 versus. 42.53%; p<.05) were also higher in the treatment group compared to control. Similarly, the viable oocytes (4.54±0.187 versus. 3.06±0.199) and the number of grade I and II oocytes per session (3.37±0.196 versus. 2.06±0.21) were higher (p<.05) in the treatment group compared with the control group. However, the nuclear maturation, cleavage, and blastocyst rate did not differ (p>.05) between the groups. Taken together, during OPU sessions, serum P4 improves oocytes' recovery and quality, whilst does not affect the in vitro developmental competence of recovered oocytes.

250 Effect of Pre- and Post-ai Nutrition on Ovarian Dynamics, Steroidogenesis, and Expression of Estrus in Beef Heifers

Abstract The objective of this study was to evaluate nutrition before and after artificial insemination (AI) on follicular dynamics, expression of estrus, and steroidogenesis. Seventy-nine beef heifers were randomly assigned to one of two dietary treatments (High;155% and Low;86% of maintenance energy) 30d prior to AI. Estrus was synchronized (PG 6-d CIDR protocol) and heifers received AI (d0) 8 to 12h following onset of estrus. On d0, heifers were randomly reassigned diets generating four Pre-x-Post-AI nutritional treatments; High-High (HH, n = 20), High-Low (HL, n = 20), Low-High (LH, n = 19), and Low-Low (LL, n = 20). Heifers remained on new diet treatments until embryo collection (flush; d7-8). Blood samples were collected daily from d-3 to d0, and on d1, 3, 5, 7 and 8 for plasma concentrations of estradiol (E2) and progesterone (P4). Dominant follicle diameter was evaluated on d-3 and d0, and CL diameter was recorded at flush. Statistical analyses were completed in SAS using the MIXED (body weight, E2, P4), GLIMMIX (expression of estrus), and GLM (ovarian parameters, interval to estrus) procedures. There were Pre-AI (P &amp;lt; 0.0001) and Post-AI by time (P &amp;lt; 0.0001) interactions on body weight; heifers in high and low treatments gained and lost weight, respectively, during both periods. Estradiol concentrations increased (P &amp;lt; 0.0001) from d-3 (2.18±0.15pg/mL) to d0 (6.05±0.04pg/mL). Pre-AI diet increased the proportion of heifers in estrus (80±6.3% vs. 59±7.9%; P = 0.05), and dominant follicle size (11.7±1.42mm vs. 10.68±1.33mm; P = 0.0016) in High vs. Low, respectively. Additionally, High Pre-AI heifers had greater P4 after AI compared to Low Pre-AI heifers (4.85±0.37ng/mL vs. 3.53±0.38ng/mL; P = 0.015). Post-AI treatment did not influence concentrations of P4 (P = 0.88). There was no effect of Pre, Post or Pre-x-Post-AI treatment on initial follicle size, follicle growth rate, E2 concentrations, interval to estrus, or CL size at flush (P &amp;gt; 0.10). In conclusion, nutrient restriction before AI negatively impacted ovarian function, steroidogenesis, and expression of estru.

Effect of exogenous administration of oxytocin on postpartum follicular dynamics, oestrous rate and ovulation in Nili-Ravi buffaloes.

Based on different surveys, dairy farmers are concerned about extensive use of exogenous oxytocin in buffaloes, which is being held responsible for reproductive problems including irregular oestrous cycle and delayed ovulation. For these concerns, effects of oxytocin injection on postpartum follicular dynamics, postpartum oestrous interval (PEI), oestrous length, the interval from onset of estrus to ovulation and blood progesterone (P4) were studied in Nili-Ravi buffaloes. For this purpose, 23 animals within 1week after calving were randomly divided into three groups: without oxytocin (CON; n=7), 10 i.u. oxytocin (LOW; n=8), 30 i.u. oxytocin - (HIGH; n=8) and used to record the PEI for the study period of 154days. At subsequent estrus, three buffaloes from each group (not served) were selected randomly to monitor two cycles for 6weeks. Transrectal ultrasonography was performed to evaluate follicular and corpus luteum (CL) development, and blood sampling was done for progesterone (P4) analysis. These results revealed that postpartum oestrous interval (PEI) decreased significantly in oxytocin-treated groups. The number of small, medium and total follicles on the left ovary was significantly higher in the HIGH group. However, an overall number of small and total follicles on both right and left ovaries was significantly higher in CON and HIGH groups. On the other hand, there was no difference in the number of follicles on the right ovary among all treatment groups. The same was true for the size of pre-ovulatory follicles, CL, P4 concentrations and oestrous cycle length. The intervals from onset of estrus to ovulation and from standing estrus to ovulation were increased considerably in the HIGH group. It is concluded that exogenous oxytocin administration resulted in the shortening of PEI but triggered a delay in ovulation. Moreover, a higher dose of oxytocin could stimulate the growth of small, medium, and total follicles in postpartum Nili-Ravi buffaloes.

Uterine Involution of Mares Supplemented with Dietary Algae-Derived Omega-3 Fatty Acids During the Peripartum Period

Different approaches have been used to improve conception rates at foal heat. Omega-3 fatty acids family and derivatives have improved reproductive efficiency in ruminants, but literature lacks studies evaluating these components on equines. The objective of this study was to evaluate effects of mare dietary supplementation with microalgae rich in docosahexaenoic acid (DHA) during peripartum on follicular dynamics and uterine involution in early post-partum. Eighteen pregnant mares, no particular breed, 410 ± 39.5 kg body weight (BW), and 7.83 ± 2.01 yr old were used. Mares were randomly assigned to control (CONT) or supplementation with microalgae rich in DHA at 0.06 g/kg BW (ALG). Treatments were supplied from 90 d prior to expected foaling date until 7 d after first ovulation. Reproductive evaluations were performed during early post-partum until 7 d after first ovulation through rectal palpation and ultrasonography of the following parameters: uterine and endometrium diameters, intrauterine fluid (IUF), uterine echogenicity, uterus tone, and follicular dynamics. Endometrial cells samples were collected to assess mRNA expression of CRP, IL-1β and AKR1C4, using RT-qPCR. Data were analyzed by mixed procedure of SAS. ALG mares had smaller uterine horns diameters and greater uterine echogenicity during post-partum in comparison with CONT. No treatment effects were detected for other characteristics evaluated, but a day effect was observed for uterine and endometrium diameter, IUF, uterine echogenicity, and transcript abundance of endometrial AKR1C4. Supplementation with DHA during peripartum may benefit uterine involution process and odds of early conception, but more studies should be performed regarding fertility.

Ovarian superstimulation with FSH in Wagyu breed bovine donors impacts folicular dynamics but does not improve the amount of embryos

The increase of the world population generates the need to raise the production of food of vegetal and animal origin. In Brazil, livestock farming has evolved, demonstrating the important role of the country in the production of food. In this way, researchers of this productive chain have been looking for technologies related to production and reproduction, above all, to the use of reproduction biotechnologies, seeking to increase the production of different bovine breeds. Thus, the objective of this research was to investigate the efficacy of ovarian FSH super stimulation in bovine Wagyu oocyte donor females on follicular dynamics and in vitro embryo production. Twelve Wagyu animals, aged 12 to 24 months, randomly distributed in a crossover design were used in two groups: Group 1= animals not stimulated with FSH and Group 2= animals stimulated with FSH. The follicular, oocyte and embryonic variables were evaluated. It was observed that ovarian overstimulation in Wagyu oocyte donor cows with FSH improved the mean and large follicles but reduced the rate of oocyte recovery and, despite the best percentage of viable oocytes, there was no improvement in the amount of embryos produced in vitro.

The Mare: A Pertinent Model for Human Assisted Reproductive Technologies?

Simple SummaryArtificial reproduction techniques (ART) are used widely in human medicine to overcome infertility, with about one in seven couples being concerned in the Western world. Due to ethical concerns, animal models are needed to develop new methodologies. Although laboratory animals are seminal in this context, they have a short lifespan and are usually fertile. Horses are long-lived domestic animals that are bred until old age, often after they have had a career being used for equestrian activities. Their reproductive functions become altered after 20 years, in a similar way to humans, although there is no menopause per se in horses. There is also a concern for rising overweight and obesity concerns in these species. In addition, embryo transfer and ART are developed to overcome infertility, as for humans. This review details similarities and differences in the reproductive cycle, ART, and fertility concerns in women and mares and discusses the opportunity of using the horse as an appropriate model for ART in humans.Although there are large differences between horses and humans for reproductive anatomy, follicular dynamics, mono-ovulation, and embryo development kinetics until the blastocyst stage are similar. In contrast to humans, however, horses are seasonal animals and do not have a menstrual cycle. Moreover, horse implantation takes place 30 days later than in humans. In terms of artificial reproduction techniques (ART), oocytes are generally matured in vitro in horses because ovarian stimulation remains inefficient. This allows the collection of oocytes without hormonal treatments. In humans, in vivo matured oocytes are collected after ovarian stimulation. Subsequently, only intra-cytoplasmic sperm injection (ICSI) is performed in horses to produce embryos, whereas both in vitro fertilization and ICSI are applied in humans. Embryos are transferred only as blastocysts in horses. In contrast, four cells to blastocyst stage embryos are transferred in humans. Embryo and oocyte cryopreservation has been mastered in humans, but not completely in horses. Finally, both species share infertility concerns due to ageing and obesity. Thus, reciprocal knowledge could be gained through the comparative study of ART and infertility treatments both in woman and mare, even though the horse could not be used as a single model for human ART.

Effects of lipopolysaccharide on beta-catenin, aromatase, and estrogen production in bovine granulosa cells in vivo and in vitro

Diseases resulting from Gram-negative bacterial infection can induce an immune response by releasing a lipopolysaccharide (LPS) endotoxin that may lead to impaired fertility in cows. To evaluate the effects of LPS on follicular dynamics in a subacute inflammatory disease state, 14 Angus heifers (BW = 413 kg±14) were blocked by weight and assigned to vehicle (n = 7) or LPS treated (n = 7) groups. Heifers received subcutaneous injections of saline (CON) or 2.0 μg/kg LPS on d 2, 5, and 8 of a select synch plus controlled internal drug release device (CIDR) follicular wave synchronization protocol. Fifty hours following CIDR withdrawal, ovaries were harvested, and follicular fluid was collected for hormone and LPS analysis. Daily blood samples were collected from d 0 to d 7. Beginning on d 8 blood samples were collected at 0, 16, 24, 32, 40, and 50 h following LPS challenge. Rectal temperatures were recorded prior to treatment and at regular intervals after each LPS challenge. Heifers treated with LPS exhibited mild (+0.5 °C) hyperthermia (P < 0.05) at 3, 4, and 8 h after the initial LPS challenge (d 2) when compared to vehicle-treated controls. Follicular fluid concentrations of estradiol (E2) increased (P = 0.04) in LPS-treated heifers compared to controls (1,595 ng/mL and 808 ng/mL±240, respectively), while follicular fluid progesterone (P4) concentrations did not differ (P = 0.27) between treatment groups. Additionally, LPS concentrations tended to be increased (P = 0.59) in dominant follicles of LPS-treated heifers, but no difference was detected (P = 0.81) in small developing follicles. To further delineate the impact of LPS on ovarian signaling pathways, a granulosa cell line (KGN) was incubated in the presence or absence of LPS (10 μg/mL) for 48 h. Cells were then collected for gene expression and protein analysis. Cells in both treatment groups expressed toll-like receptor 4, myeloid differentiation factor-2 receptor, and CD-14 complex genes required for LPS signaling. Cells treated with LPS exhibited decreased mRNA expression of aromatase (P = 0.03) and beta-catenin (P = 0.02). However, no change (P > 0.10) was detected in abundance of total beta-catenin protein or beta-catenin phosphorylated isoforms at serine 552 or 675. Based on results from this in vivo experiment, these investigators concluded that low doses of LPS can alter E2 concentrations and this effect may be modulated in part through beta-catenin regulation of aromatase transcription.

Studies on Ovarian Follicular Dynamics and Steroid Profiles in Sahiwal Cattle

Background: Understanding of ovarian follicular dynamics and endocrine regulation is essential to design and use interventions to optimize reproductive efficiency. Bos indicus and Bos taurus cows have some differences in their follicular dynamics and ovarian steroidal hormones. The present study was planned to understand follicular dynamics vis-a-vis ovarian steroids profiles in Sahiwal cattle. Methods: The study was conducted in normal cyclic, pluriparous, non-lactating Sahiwal cows (n=7). Trans-rectal ultrasonography was performed to record the location, size, number of follicles and size of corpus luteum (CL) from the beginning, till the end of experiment. The dataset was used to characterize the follicular wave emergence, growth, regression, time of selection and ovulation of the dominant follicle in each cow. Result: Three Sahiwal cows showed two wave and four cows showed three wave estrous cycles with mean interovulatory length of 20.33±0.33 and 22.50±0.28 days, respectively. No significant difference was recorded in the maximum diameter of corpus luteum, P4 and E2 plasma levels between 2-wave and 3-wave estrous cycles. The peak progesterone values of 6.00±0.91 and 6.2±1.2 ng/ml and peak estradiol values of 15.83±0.60 and 14.31±0.44 ng/ml were recorded in 2-wave and 3-wave estrous cycle, respectively. The results showed that Sahiwal cows had 2-wave and 3-wave estrous cycle and the 3-wave estrous cycle had comparatively longer inter-ovulatory period and smaller maximum diameter of second wave dominant follicle than 2-wave estrous cycle.