Total Number Of Follicles Research Articles

Causes and consequences of the variation in the number of ovarian follicles in cattle

Summary. In cattle we have noted that the antral follicle count (AFC, follicles ≥3 mm in diameter) varies greatly among animals (from 5 to 50), is repeatable within animals, and is highly correlated with the total number of healthy follicles in ovaries. Also, animals with low AFC have higher serum concentrations of FSH and LH, but lower concentrations of Anti-Mullerian Hormone, progesterone and androgens than animals with high AFC. We have investigated the effect of maternal environment during gestation on their offspring AFC by restricting maternal nutrition to 60% of maintenance requirements (compared with 100% in controls) during the first third of gestation. Calves born to nutritionally restricted mothers had 60% lower AFC compared with calves born to mothers fed control diets. In other studies we have evidence to indicate that fertility may be compromised in animals with low AFC due to effects on oocytes, progesterone and the endometrium compared with animals with high AFC. To examine this directly we assessed AFC in post-partum dairy cows and found that cows with a high AFC had higher pregnancy rates, shorter calving to conception intervals and received fewer services during the breeding season compared with cows with a low AFC. In addition, the high variation in follicle numbers in adults may not only be reflective of reproductive disorders and suboptimal fertility, but there is evidence to indicate that it may be associated with alterations in the function of other non-reproductive systems (e.g. cardiovascular) that may have profound effects on the animal's health and welfare.

Inhibin and oestradiol in the control of FSH secretion in the sheep

In the sheep both FSH and LH are necessary for development of large antral follicles. The secretion of FSH is controlled by the negative feedback effect of two ovarian hormones, oestradiol and inhibin, acting at the level of the anterior pituitary. Both are derived from the granulosa cells of large antral follicles which are present in sheep ovaries throughout the oestrous cycle. FSH stimulates growth and mitosis and so the fully differentiated granulosa cells of the large preovulatory follicles acquire receptors for LH, have maximal aromatase activity and produce large amounts of inhibin. The number of these large antral follicles (oestrogenic) which have the potential for ovulation corresponds to the ovulation rate specific for each particular breed of sheep. Over 90% of the oestradiol secreted by the ovaries is derived from these chosen follicles. In contrast, inhibin (and androstenedione) is also secreted by large antral follicles which have lost or not yet acquired maximal aromatase activity. The secretion of oestradiol by the preovulatory follicle(s) is dependent on the supply of androgen precursors produced by the theca which is stimulated by LH. When the concentration of progesterone falls at the end of the luteal phase the increased secretion of LH stimulates the progressive increase in secretion of oestradiol which occurs during the follicular phase. At this stage of the cycle there is a modest inconsistent rise in inhibin, the secretion of which is not stimulated by LH but is related to the increase in the number of large antral follicles. It is suggested that inhibin with its long half-life sets the overall level of negative feedback while oestradiol is responsible for the day-to-day fluctuations in the concentration of FSH which determines the number of ovulatory follicles. This dual control of FSH is adapted to monitor both the total number of large antral follicles in the ovaries (inhibin) and the number which are selected for ovulation (oestradiol).

Effect of oral administration of pioglitazone on follicular dynamics in Holstein dairy cows

This study investigated the effects of oral administration of pioglitazone (PGT), a specific and synthetic ligand of peroxisome proliferator-activated receptors gamma (PPARγ), on follicular dynamics and corpus luteum (CL) functionality in dairy cows. Cows exhibiting strong signs of estrus after 2 injections of PGF2α (given14 d apart) at d 30 postpartum (n = 28) were allotted to four groups (n = 7 cows/treatment) and orally received 6 mg PGT/kg body weight/day according to the following protocol: no PGT (control); PGT for 14 d from 7 d before expected estrus (10 d after 1st injection of PGF2α) to 7 d after observed estrus (PGT14); PGT for 21 d after observed estrus (PGT21); and PGT for 28 d, 7 d before expected estrus to 21 d after observed estrus (PGT28). During the first follicular wave, number of follicles (total and small) increased in PGT14 and PGT28 cows compared to the control group (P < 0.05). During the ovulatory wave, number of total and small follicles increased in PGT28 (P < 0.05) and PGT21 (P < 0.10) compared with PGT14 and control cows. Size of the largest follicle at first wave was greater in PGT28 (P < 0.05), PGT14 (P < 0.05) and PGT21 (P < 0.10) compared to the control cows. Maximal size of the ovulatory follicle was greater in PGT28 (P < 0.05) and PGT21 (P < 0.10) groups compared to the control group. Growth rate of the largest follicle at first wave was higher (P < 0.05) in PGT-treated cows, while growth rate of the ovulatory wave was higher in PGT28 and PGT21 groups, leading to shorter days from luteolysis to ovulation. Pioglitazone administration did not affect CL size, but increased progesterone (P4) concentration. The PGT14 and PGT28 cows had higher maximal plasma P4 concentration and shorter intervals to reach maximal plasma P4 compared to the control group. In conclusion, oral administration of PGT had some positive effects on follicular development and circulating P4 levels which may be conducive to better reproductive performance.

Body mass index and Ovarian response in an In-Vitro Fertilization Cycle

Background: In-vitro fertilization (IVF) has established its place in the management of infertility cases that has defied conventional treatment. A successful In-vitro fertilization outcome is based on a good ovarian response to gonadotrophin stimulation. Body mass index has been identified as one factor that can affect ovarian response however; there is no consensus about its impact outcome.Objectives: To determine the effect of body mass index (BMI) on ovarian response in an Invitro fertilization cycle.Method: A Prospective cohort study to determine the response to Controlled Ovarian Hyperstimulation (COH) in women with normal and abnormal BMI in an IVF cycle. One hundred and four (104) consenting women undergoing non-donor IVF cycles in a university teaching hospital were recruited for the study. Selected participants had their body mass indices (BMI) assessed and classified into 3 groups: Normal weight (18.5-24.9kg/m2), overweight (25-29.9kg/m2) and obese (≥30kg/m2). There were no underweight women-BMI &lt;18.5kg/m2. All the participants had Controlled Ovarian Hyperstimulation with the long agonist protocol.Selected participants had follicular count assessed using a transvaginal sonography on day 3, 5, 10 and at retrieval. The effect of ovarian response with the IVF treatment assessed based on total number of follicles and oocyte yield on the day of retrieval.Results: Female participants with normal weight women were thirty-six (34.6%), overweight were forty (38.5%) and obese, twenty-eight (26.9%). There was no statistical significant difference when the ages of women at presentation, total gonadotrophins requirements, duration of stimulation and number of oocytes retrieved were compared in the three groups.Conclusion: Increase in body mass index does not appear to have an adverse effect on ovarian response in an In-vitro fertilization cycle._________________________________________________________________________Keywords: body mass index, infertility, ovarian stimulation, gonadotrophins, IVF cycle.

Effects of follicle-stimulating hormone followed by gonadotropin-releasing hormone on embryo production by ovum pick-up and in vitro fertilization in the river buffalo (Bubalus bubalis).

In this study, we examined the effects of superstimulation using follicle‐stimulating hormone (FSH) followed by gonadotropin‐releasing hormone (GnRH) on buffalo embryo production by ultrasound‐guided ovum pick‐up (OPU) and in vitro fertilization (IVF). Nine Murrah buffaloes were subjected to OPU‐IVF without superstimulation (control). The morphologies of the oocytes collected were evaluated, and oocytes were then submitted to in vitro maturation (IVM). Two days after OPU, same nine buffaloes were treated with twice‐daily injections of FSH for 3 days for superstimulation followed by a GnRH injection. Oocytes were collected by OPU 23–24 hr after the GnRH injection and submitted to IVM (the superstimulated group). The total number of follicles, number of follicles with a diameter > 8 mm, and number of oocytes surrounded by multi‐layered cumulus cells were higher in the superstimulated group than in the control group (p ≤ 0.05). After IVF, the percentages of cleavage and development to blastocysts were higher in the superstimulated group than in the control group (p < 0.05). In conclusion, superstimulation improved the quality of oocytes and the embryo productivity of OPU‐IVF in river buffaloes.

Impact of uteroplacental insufficiency on ovarian follicular pool in the rat

BackgroundA low oxygen supply to the fetus causes intrauterine growth restriction and can affect gonadal development of the offspring, having a potential impact on fertility. We investigated histology and gene expression in the postnatal rat ovary after fetal hypoxia induced by uterine artery ligation.MethodsSprague-Dawley rats underwent uterine artery ligation at day 19 of gestation. Offspring were sacrificed at 5, 20 and 40 days post-partum. Follicles were counted and classified in hematoxylin-eosin stained sections. Gene expression of 90 genes was analyzed by TaqMan® Low Density Array.ResultsA significantly lower number of total and primordial follicles was detected in 20 days post-partum intrauterine growth restricted animals. Follicle density was not different at 40 days post-partum, suggesting that compensatory mechanisms occurred during the pre-pubertal window. Uterine artery ligation modified the expression of 24 genes involved in different cellular functions, among which proliferation, apoptosis and metabolism.ConclusionOvarian follicle pool was affected by fetal hypoxia in early life, but this effect did not persist in puberty. Genes involved in cellular processes were affected at all ages, potentially implying long-term genetic alterations. Further analyses are needed to elucidate later effects of fetal hypoxia on ovarian function and fertility.

Effect of photoperiod on follicular IGF-1 and oocyte quality independently of metabolic status in buffalo heifers

The aim of this study was to determine whether buffalo heifers maintained at a constant live weight (LW) and body condition score (BCS) by a restricted diet show changes in reproductive function in response to changes in day length. Heifers were fed a low energy diet (86% daily intake recommended for the species) throughout the study, which involved an 8-week period of decreasing day length and a 7-week period of increasing day length. Weekly, LW, BCS and ovarian follicular/oocyte population were assessed. Blood and follicular fluid samples were taken at the time of ovum pick-up for hormone analyses and at 30-day intervals to measure metabolic substrates. Buffalo heifers maintained LW and BCS throughout the study and blood concentrations of the main metabolic hormones, such as GH, insulin, IGF-1 and thyroid hormones were not influenced by photoperiod. Likewise, there were no differences in metabolic substrates such as glucose, NEFA, triglycerides and HDL cholesterol, suggesting that the metabolic condition remained essentially the same. During increasing day length periods, the number of total follicles (8.4 ± 0.5 vs. 11.9 ± 0.6; p < .01), cumulus-oocyte complexes (2.2 ± 0.3 vs 4.3 ± 0.5; p < .01) and superior quality Grade A + B COCs (0.2 ± 0.1 vs 1.1 ± 0.2; p < .01) decreased. These features were associated to reduced oestradiol levels both in plasma (3.9 ± 0.4 vs 9.2 ± 0.5 pg/mL; p < .01) and follicular fluid (49.7 ± 12.9 vs. 109.1 ± 25.8 ng/mL; p = .05), as well as to reduced intrafollicular IGF-1 levels (45.5 ± 4.0 vs. 58.5 ± 3.9 ng/mL; p < .05). These findings suggest that the ovarian function in buffalo heifers is influenced by photoperiod independent of nutritional and metabolic status.HighlightsMetabolic profile, LW and BCS do not change in buffalo heifers on a restricted dietPhotoperiod affects ovarian function in heifers regardless of metabolic statusFollicular and oocyte population decrease in increasing day length periodsOestradiol and IGF-1 levels in follicular fluid are affected by photoperiod

Short Letrozole Therapy Vs Extended (Long) Letrozole Therapy for Induction of Ovulation in Women with Polycystic Ovary Syndrome

Background: polycystic ovary syndrome (PCOS) is the major cause of anovulatory infertility. Letrozole and other aromatase inhibitors (AIs) have been introduced as a new treatment option could challenge clomiphene citrate (CC) for ovulation induction in this group. Objective: to compare the efficacy of short vs extended courses of letrozole therapy for ovulation induction in PCOS women. Patients and Methods: this is prospective randomized comparative study done on 60 patients attended El-Hussein Hospital and previously diagnosed as having PCOS, based on the revised Rotterdam criteria (2003). Patients were divided randomly into 2 treatment groups, group I, short letrozole therapy (30 patients, received 5mg of letrozole from 1st to 5th day of menstrual bleeding) and group II, long letrozole therapy (30 patients, received 2.5mg of letrozole from 1st to 10th day of menstrual bleeding). All patients of both groups monitored by trans-vaginal ultrasound on day 10 of the cycles. HCG injection (5,000 IU IM) was given when at least one follicle measured ≥ 18mm. Intercourse were advised for 24-36h after HCG injection. Serum HCG and trans-vaginal ultrasound were done 2 weeks after HCG injection in the absence of menstruation for diagnosis of pregnancy. Results: in long letrozole therapy, total number of follicles, mean number of mature follicles, endometrial thickness, percentage of patients ovulated after treatment, occurrence of pregnancy were greater when compared to results of short letrozole therapy. Conclusion: long letrozole therapy can produce more mature follicles and subsequently more pregnancies than short letrozole therapy.

Vitamin C restores ovarian follicular reservation in a mouse model of aging.

Ovarian aging is related to the reduction of oocyte quality and ovarian follicles reservation leading to infertility. Vitamin C is a natural antioxidant which may counteract with adverse effects of aging in the ovary. The aim of this study was to evaluate the possible effect of vitamin C on NMRI mice ovarian aging according to the stereological study. In this experimental study, 36 adult female mice (25–30 g) were divided into two groups: control and vitamin C. Vitamin C (150 mg/kg/day) were administered by oral gavage for 33 weeks. Six animals of each group were sacrificed on week 8, 12, and 33, and right ovary samples were extracted for stereology analysis. Our data showed that the total volume of ovary, cortex, medulla and corpus luteum were significantly increased in vitamin C group in comparison to the control groups (P≤0.05). In addition, the total number of primordial, primary, secondary, and antral follicles as well as granulosa cells were improved in vitamin C group in compared to the control groups (P≤0.05). No significant difference was observed in total volume of oocytes in antral follicles between control and vitamin C groups. Our data showed that vitamin C could notably compensate undesirable effects of ovarian aging in a mouse model.

Assessment of Safe Donor Zone of Scalp and Beard for Follicular Unit Extraction in Indian Men: A Study of 580 Cases.

Context:Hair restoration surgery for androgenetic alopecia (AGA) essentially involves various forms of hair transplantation. There is paucity of studies assessing donor area in Indian men and also no simplified guidelines are available for the safe donor area for follicular unit extraction (FUE). Our study is an attempt to study the donor area in Indian men.Aims:To assess the density of follicular units (FUs) in the donor area, that is, both scalp and beard in Indian men, and to propose simplified guidelines for FUE.Materials and Methods:The study design was cross-sectional and was carried out for 2 years. All the consenting male patients with male pattern hair loss Hamilton Norwood grading III or more who consulted for a hair restoration surgery were recruited. FU density was assessed in the donor area of scalp by drawing a rectangle with its lower border being a straight line joining two points, which are 27–28mm from the line drawn perpendicular to tragus, passing through external occipital protuberance. Three squares of area 1cm2 were drawn within the rectangle. Average of the FUs and follicles in the three squares was calculated to obtain mean density in the donor area of the scalp. Total number of FU was assessed, considering 25% extraction, total average number of extractable follicles was assessed. Total donor area was divided into three areas (areas 1, 2, and 3) and average number of extractable FU was assessed in each. Donor area in the beard, below the jawline, was divided into a triangle and rectangle. Average number of FU, total number of extractable FU was calculated similarly.Results:A total of 580 male patients were recruited in the study. Mean FU density in the scalp and beard was 78.2/cm2 and 49.7/cm2, respectively. The total available number of FUs for extraction in the areas 1, 2, and 3 and beard considering 25% extraction was 2064, 3097, 3612, and 824, respectively. We propose three types of donor areas in the scalp, namely, limited, standard, and extended donor area.

A study of the histopathological features of alopecias on transverse sections of scalp biopsies

Background:Transverse sections of scalp biopsies are performed for the assessment of alopecias and are considered advantageous over vertical sections.Aim:The aim was to study the histopathological features of alopecias on transverse sections of scalp biopsies.Methods:It was a descriptive study. Clinically confirmed cases of noncicatricial and cicatricial alopecias were subjected to 4 mm scalp biopsies, which were sectioned transversely and analyzed. Biopsies obtained from occipital region of androgenetic alopecia (AGA) cases were taken as controls.Results:Biopsies from 41 cases were assessed, including male and female AGA, alopecia areata (AA), trichotillomania, lichen planopilaris (LPP), discoid lupus erythematosus (DLE), and folliculitis decalvans (FD). Normal scalp (control) biopsies showed the median total number of hair follicles of 35 (32–37), anagen:telogen/catagen ratio of 17.5 (16.5–31), and terminal:vellus ratio of 15 (10.7–17.5). In AGA and AA, miniaturization and shift toward telogen and catagen hair were consistently observed. Peribulbar inflammation was seen in two-third of AA. Trichotillomania showed increased catagen hair and numerous pigment casts. In DLE, besides perifollicular inflammation, prominent peri-arrector pili and peri-eccrine inflammation were observed. Type of inflammatory infiltrate was similar in DLE and LPP (lymphocytic), whereas FD showed neutrophilic and plasma cell infiltrate, both around follicles and interstitially. Basal cell damage in the follicles and pigment incontinence were seen in majority of DLE and LPP patients. DLE also showed basement membrane thickening, mucin deposition, and telangiectasia. Reduction/absence of sebaceous glands and perifollicular fibrosis were observed in almost all cicatricial alopecias.Conclusion:Transverse sectioning may be a useful tool in the diagnosis of alopecias.

Assessment of reflectance confocal microscopy for non-invasive selection of optimal ovarian cortex fragments for autotransplantation.

Can reflectance confocal microscopy (RCM) be used to determine follicle density in human ovarian cortex fragments that are intended for fertility restoration? RCM was used on living cortex tissue fragments derived from five bovine ovaries and 13 human ovaries. All tissue fragments were cryopreserved and thawed before RCM analysis. Follicle numbers and distribution were determined by RCM and histology. Before and after RCM, general tissue viability and follicle integrity were assessed by a glucose uptake assay and neutral red staining, respectively. RCM can detect all stages of follicle development in living ovarian tissue to a maximum depth of 250 µm. In bovine tissue, all follicles were located within this 0-250 µm range. In human ovarian tissue, follicles were also present below the 250 µm RCM threshold, implying that only a percentage of the total number of follicles could be detected with RCM. The percentage of follicles detected by RCM appeared to be age dependent. The RCM procedure did not affect the glucose uptake by the tissue, whereas neutral red staining indicated a high level of follicle survival. In this proof of concept study, we have shown that RCM is a promising technique to determine the density of follicles ex vivo in living human ovarian cortex fragments, apparently without compromising the vitality of the tissue. Safety studies and further optimization of the RCM technique with a focus on increasing the penetration depth are required before clinical use of RCM.

Stimulatory effects of fish oil and vitamin E on ovarian function of laying hen

In recent years, there has been an increase in the consumption of omega-6 fatty acids (FAs) over omega-3. Moreover, the antioxidant nature of oil supplemented diet in human and animal studies is not clear. In this work, the n-3 effects on hen ovarian cells, ovarian dynamics, serum FA profiles and expression level of apoptosis genes in the presence or absence of vitamin E were studied. Sixty-eight laying hens were randomly divided into four groups and the groups were fed the following diets: basal diet +1.5% sunflower oil (control; C); basal diet +1.5% sunflower oil +1.1 U alpha-tocopherol/hen/day (E); basal diet +1.5% fish oil +1.1 U alpha-tocopherol/hen/day (n-3 + E); and basal diet +1.5% fish oil (n-3). In E, n-3 + E and n-3, small yellow follicle numbers were higher than control (p < .05). Large yellow follicle numbers as well as total numbers of follicles of n-3 + E and n-3 were higher (p < .05) than control and E. The rate of ovulation was significantly (p < .05) higher in n-3 (96%) compared to the control (89%). The proportion of intact primordial follicles in n-3 and n-3 + E was significantly more than control and E (p < .01). The expression of Bak (Bcl2 antagonist/killer) was significantly (p < .05) higher in control than the other groups. There was significant reduction in expression of Cas9 in n-3 and E compared to the control and n-3 + E (p < .05). While omega-3 and vitamin E can protect ovarian cells from apoptotic death, omega-3 source could have promoting roles on follicular development and ovulation rate, being independent of dietary vitamin E.HighlightsDespite the perceived importance of supplementing antioxidant with fatty acids, our results indicated that alone low level of n-3 has a great influence on ovary performance.

Follicle populations and gene expression profiles of Nelore and Angus heifers with low and high ovarian follicle counts.

The number of antral follicles counted (AFC) by ultrasound is associated with fertility in cattle. Cows with higher follicle count (HFC) have higher performance in reproductive-assisted technologies than cows with lower follicle count (LFC). In this study, we aimed to define the preantral follicle count by histology and to identify differentially expressed genes (DEGs) using a microarray in Nelore and Angus heifers with HFC and LFC. The ovaries of each animal were scanned with an ultrasound device 12 to 24 hr after estrus. The groups were formed based on the average number of total follicles (≥3 mm) counted in each breed consistently ± the standard deviation. For the histological analysis, preantral follicles were counted and classified under a stereo microscope, and follicle density was determined. Microarray analysis was performed on pools of three follicles dissected from the ovaries of 15 Nelore (6 HFC and 9 LFC) and 17 Angus heifers (9 HFC and 8 LFC). Angus heifers have increased total and primordial follicle density. Nelore heifers have increased antral follicle count. Different patterns of gene expression regulate follicle recruitment and development in Angus and Nelore heifers and may be associated with the different follicle densities observed in Angus versus Nelore heifers. Furthermore, HFC heifers presented increased expression of genes associated with cellular development and metabolism.

135 Use and dose of porcine follicle-stimulating hormone for ovarian superstimulation prior to ovum pickup and in vitro embryo production in pregnant Holstein heifers

The benefit of superstimulation with exogenous FSH before ovum pickup for in vitro embryo production has been the subject of significant controversy. In addition, there is limited information on different dose regimens. Thus, the objective of the present study was to evaluate the effect of dose of porcine (p)-FSH during superstimulation before ovum pickup (OPU) on in vitro embryo production in pregnant heifers. Pregnant Holstein heifers (n=36) were assigned to a complete 3×3 crossover design. Three treatment groups were evaluated as follows: p-FSH 0mg (FSH0), p-FSH 160mg (FSH160) and p-FSH 300mg (FSH300). Three sessions of OPU were performed on each animal at 48, 62 and 76 days of gestation, with a washout interval between sessions of 14 days. Follicular wave emergence was synchronized by dominant follicle removal. Heifers in the FSH0 group received no further treatment, whereas the remaining groups received a total of 4 injections 12h apart as follows: FSH160 (48.0, 42.7, 37.3 and 32.0mg) or FSH300 (90.0, 80.0, 70.0 and 60.0mg), beginning 36h after dominant follicle removal. Ovum pickup was performed in all heifers 40h after the last p-FSH injection. Heifers were subjected to OPU for oocyte recovery, and number of follicles was determined. Recovered oocytes were processed and in vitro embryo production performed. Differences between treatment groups were evaluated by generalized linear mixed models. Data are presented (Table 1) as mean±standard error of the mean. There was no effect of days in gestation for any of the outcomes evaluated (P&amp;gt;0.05). Follicle numbers at the time of oocyte recovery were different (P&amp;lt;0.01) between groups. Heifers in the FSH300 group had a greater (P&amp;lt;0.05) number of medium, large and total follicles than heifers in the FSH0 group, whereas heifers in the FSH160 were intermediate. Total number of recovered, viable and cleaved oocytes were greater (P&amp;lt;0.01) in FSH300- than in FSH160- and FSH0-treated heifers. Cleavage rate and blastocyst development rate were not different (P&amp;gt;0.10) between groups. The number of grade 1 and 2 blastocysts was greater in FSH300- than in FSH160- and FSH0-treated heifers (P&amp;lt;0.03). In summary, the use of 300mg of p-FSH before OPU in pregnant heifers increases the number of follicles, oocytes and blastocysts produced per heifer with no detrimental effect on oocyte competence. Table 1.Ovum pickup and in vitro embryo production in pregnant heifers treated with different doses of porcine FSH

133 Test of minimum-intervention protocols for optimizing in vitro embryo production in bison

The study was done to determine whether minimal handling protocols for ovarian synchronization and ovarian superstimulation may be used to increase in vitro embryo production in bison. Ultrasound-guided cumulus-oocyte complex (COC) collection was done in a group of bison (n=23; random start) during the anovulatory season to synchronize new follicular wave emergence. The COC were classified morphologically (compact-good and -regular, expanded, denuded, degenerate) but not processed further. At the time of COC collection (Day 0), bison were assigned randomly to 3 groups and given 5mL of saline IM (non-superstimulated controls; n=11), 10 Armour units of pFSH (Antrin R10, Kyoritsu Seiyaku Corp., Tokyo, Japan) in 5mL of saline IM once per day from Day 0 to 2 (regular FSH; n=5), or 30 armour units of a sustained-release form of pFSH (Antrin R10Al, Kyoritsu Seiyaku Corp.) in 5mL of saline SC on Day 0 (long-acting FSH; n=7). On Day 4, a second COC collection was performed. Only compact COC were processed. The COC were matured in vitro for 25 to 28h at 38.8°C, fertilized (2×106 sperm mL−1) and co-incubated at 38.8°C in 5% CO2 for 18h. Presumptive zygotes were denuded and cultured at 38.8°C in 5% O2, 5% CO2 and 90% N2. Nominal data were compared by t-test and analysis of variance. Binomial data were compared among groups by chi-squared. There was no difference between the first (random) COC collection (n=23) and second collection (n=11 non-superstimulated controls) in the total number of follicles detected, but the distribution among size categories (3-4, 4-8, and &amp;gt;8mm) differed, i.e. fewer in the 3 to 4mm category at the time of the second COC collection (12.2±1.0v. 8.1±1.4; P&amp;lt;0.05). In the nonstimulated control group, there were no differences between the first and second COC collections in the number of follicles aspirated (12.7±1.0v. 10.4±1.5), number of COC collected (7.7±0.9v. 5.3±1.3), or in the categorical distribution of COC. At the second COC collection, the number of follicles in the &amp;gt;8mm category was greater in the regular FSH group than in the control or long-acting FSH groups (2.8±0.5v. 1.1±0.3, and 1.9±0.4, respectively; P&amp;lt;0.05), but no differences were detected in the number of follicles aspirated, COC collected, or in the categorical distribution of COC. The cleavage rate (of total oocytes submitted to in vitro maturation), recorded 2 days after IVF, was higher in the control group than in either the regular FSH or long-acting FSH groups [25/35 (71.4%), 7/28 (25.0%), 8/35 (22.8%); P&amp;lt;0.0001]. The freezable embryo production rate, recorded 7 days after IVF, was greater in the control group than in the regular FSH or long-acting FSH groups [19/35 (54.3%), 5/28 (17.9%), 5/35 (14.3%); P&amp;lt;0.01]. In conclusion, minimal-handling interventions used in the present study to increase embryo production in bison were not effective, likely as a result of the timing, frequency, and duration of superstimulation. A random start resulted in greater COC collection than collection 4 days after ovarian synchronization, and embryo production rates were greater in non-superstimulated bison. This work was supported by Parks Canada and Saskatchewan ADF. Antrin products donated by Kyoritsu Seiyaku Corp., Japan.

Combined Letrozole and Clomiphene versus Letrozole with low dose gonadotropin protocol for ovulation induction in infertile clomiphene-resistant women with polycystic ovary syndrome: Comparative study

Objectives: To compare the effect of combined clomiphene citrate (100mg) and Letrozole (5mg) versus Letrozole (5mg) followed by gonadotropin injection for one cycle in CC resistant PCOs patients.Design: Prospective randomized trial.Setting: Infertility Outpatient Clinic and Cytogenetic and Endoscopy Unit of Obstetrics and Gynecology Department of Zagazig University Hospitals.Study group: About 212 PCOs patients with CC resistance (anovulatory response with CC at a dose of 100-150 mg for 3 cycles) were divided into two groups (106 in each group).Intervention: A studied group was randomly divided into two groups. Group A received combined Letrozole 5mg (2 tablets) in the morning and clomid 100 mg (2 tablets) in the evening from the 3rd day of menses for 5 days, group B received Letrozole 5 mg (2 tablets) from the 3rd day of menses for 5 days followed by human menopausal gonadotropin (hMG) injection every other day from cycle day 7. Moreover, ultrasound folliculometry started on cycle day 10 then the dose was adjusted according to follicular size. Both groups received treatment regimen for only one cycle.Results: There was higher ovulation rate (85.8%) in letrozole plus hmG group than in the other group receiving letrozole and CC (80.2%), but with no statistical significant difference (P -value =0.65). But, there was statistical significant difference between both groups regarding stimulation characters including total number of mature follicles (≥18 mm) 2.1 ± 1.01 in combined letrozole and CC was greater in group A than in group B (1.66 ± 0.98) with (P - value = 0.009). Stimulation days were longer in group B (18.2 ± 2.5) than in group A (16.2 ± 2.1). Mean total E2 on the day of hCG administration was significantly higher among patients in the clomiphene-letrozole group as compared with that in the other group of letrozole with hmG ( 650 ± 190.3 and 540 ± 180.5, respectively). Also, higher day 21 serum progesterone in group A (11 ± 1.1) than (9.5 ± 0.9) in group B. But, there was no statistical significant difference in mean endometrial thickness in both groups (A and B) (9.6 ± 1.7 versus 9.8 ± 1.5mm, respectively) (P-value = 0.41). The pregnancy rate did not show any statistical significant difference in both groups (P-value = 0.45) ; 32 patients out of 106 (30.1%) in group A compared with 27 patients (25.4%) in group B. Four cases were aborted in group A compared with 3 cases in group B. Four cases had twin pregnancy in group A with only 3 cases in group B. Also, there was no significant difference between both groups regarding ovarian hyperstimulation ; as there were 2 cases with combined clomid and letrozole group and one case only in letrozole plus hmG group.Conclusion: This study showed that lerozole is an effective medication when combined with CC or sequentially with hmG injection in improving the ovulation and pregnancy rate in CC-resistant PCOs patients with higher results when combined with hmG injection, but did not reach statistical significance.

Involvement of Quebracho tannins in dietalters productive and reproductive efficiency of postpartum buffalo cows.

This study was conducted to investigate the effects of 10 weeks supplementation of Quebracho tannins (QT; 0 [control], 100 [QT100] or 200 g/[cow·d] [QT200]) to 30 multiparous postpartum buffalo cows (10 cows per group) on milk yield and composition, blood metabolites and reproductive performance. Supplementation of QT100 had no significant effect on milk yield, whereas QT200 decreased (P < 0.05) this trait. Compared with the control group, both QT levels decreased (P < 0.05) fat-corrected milk (FCM) yield, but no significant effects were found on percentages of milk fat and protein. Contrariwise, yields of milk fat, lactose and milk protein were decreased (P < 0.05) when QT200 was supplemented. The solids nonfat (SNF) percentage and yield were decreased (P < 0.05) with QT100 supplementation. Moreover, QT tended to numerically reduce total number of ovarian follicles, number of small follicles, peripheral progesterone concentration and conception rate. Supplementation of QT200 numerically increased number of large follicles, mean diameter of large follicle, number and diameters of corpora lutea. The inclusion of QT200 shortened days open (DO) and decreased number of services per conception. Contrariwise, QT did not show significant effects on serum total protein, albumin, globulin, glucose, cholesterol and triglycerides concentrations. Supplementation of QT100 caused an increase (P < 0.05) of serum urea compared with that in control and QT200 groups. Generally, QT decreased (P < 0.05) serum creatinine concentration. Therefore, the supplementation of a commercial QT to early lactating Egyptian buffalo cows displayed negative consequences on their productive and reproductive performances.

The potential reproductive toxicity of tannery effluent to the estrous cycle and ovarian follicular dynamics of female Swiss mice.

Although the toxic effects of tannery effluent (TE) on tanning-industry workers have been reported in many studies, its effects on females' reproductive system are unknown. We aimed at evaluating the effects of direct contact with TE on the "emotional" status, estrous cycle (during 15 consecutive exposure days), and ovarian follicular dynamics of female Swiss mice at the end of the experiment to broaden the knowledge about the toxicity of this pollutant. The herein adopted exposure protocol simulated tanning-industry workers' exposure to TE. The test animals were subjected to 45 exposure days, for 1h a day, 5days a week (from Monday to Friday). Based on the collected data, female mice exposed to TE recorded high anxiety index in the elevated plus maze test, although we did not observe changes in their estrous cycle. The smaller total and specific number of ovarian follicles (types 1 to 6) and the higher frequency of degenerating follicles (atresic) in female mice exposed to TE marked the folliculogenesis reduction in them. Therefore, our study was the first to provide evidences that the exposure to TE can cause reproduction issues in female mice, as well as the first experimental insight about the impact of unhealthy work activities in tanning industries on women's reproductive system.

Extended Letrozole Therapy for Ovulation Induction in clomiphene resistant Women with Polycystic Ovary Syndrome

Objective: This study aimed to evaluate the efficacy of short and extended long course of letrozole therapy for ovulation induction in clomiphene resistant women with polycystic ovary syndrome. Patients: One hundred infertile women were selected from the patients attending the outpatient clinic of Manshiet El-Bakry general hospital. All patients were diagnosed as having anovulation due to polycystic ovary syndrome (PCOS). Interventions: Patients were randomly allocated to treatment with either long letrozole group took 2.5 mg of letrozole daily starting day 1 of spontaneous or progesterone inducing menstrual bleeding for 10 days (50 patients, up to 3 cycles) or short letrozole group took 5 mg of letrozole daily starting day 1 of spontaneous or progesterone inducing menstrual bleeding for 5 days (50 patients, up to 3 cycles). Results:The number of ovulating patients was greater in the long letrozole group (74% vs. 56%), but without statistical differences. The total number of follicles during stimulation was insignificantly greater in the long letrozole group (8.2 vs. 8.17). The numbers of follicles ≥18 mm were significantly greater in the long letrozole group. Pregnancy occurred in 7 in the short group (14%) and 12 of (24%) in the long letrozole group, and the difference was statistically insignificant. Conclusion: The long letrozole protocol (10 days) can produce more mature follicles and subsequently more pregnancies than the short letrozole therapy (5 days).