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SummaryThe goal of this study was to determine the distribution of preantral follicles in bovine ovaries. Follicular distribution in the ovaries (n = 12) was evaluated in the region of the greater curvature of the ovary (GCO) and the region close to the ovarian pedicle (OP) of Bos taurus indicus heifers of the Nelore breed. Two fragments were obtained from each region of the ovary (GCO and OP). The mean weight of the ovaries was 4.04 ± 0.32 g. The mean antral follicle count (AFC) was 54.58 ± 3.55 follicles (minimum and maximum variation of 30 and 71 follicles, respectively). In total, 1123 follicles were visualized in the region of the GCO; 949 (84.5%) of them were primordial follicles and 174 (15.5%) were developing follicles. The region close to the OP contained 1454 follicles, of which 1266 (87%) were primordial follicles and 44 (12.9%) were developing follicles. The OP region showed a higher proportion of intact follicles in the primordial (P < 0.0001) and primary (P = 0.042) stages compared with the GCO region. The proportion of secondary follicles was similar in the OP and GCO regions. The ovaries of two bovine females (16%; 2/12) contained multi-oocytes follicles, which were characterized as primary follicles. Therefore, the distribution of preantral follicles in the bovine ovary was heterogeneous, with the region close to the OP containing a greater number of preantral follicles compared with the GCO region (P < 0.05).

The aim of this study was to evaluate follicular development, morphological integrity, and antioxidant potential of preantral ovarian follicles from Bos taurus indicus females grown in vitro with alpha-lipoic acid. Ovaries (n = 24) of Bos taurus indicus (n = 12) females were collected during slaughter and fragmented. A randomly obtained fragment from each pair of ovaries was fixed in Bouin (non-cultivated control; D0). These fragments were intended for classical histology (morphology and evaluation of follicular growth), and a fragment from each pair of ovaries was frozen at -80°C (non-cultivated control; D0), and assigned for analysis of oxidative stress [thiobarbituric acid reactive substances (TBARS), nitroblue tetrazolium (NBT), and ferric reducing antioxidant power (FRAP)]. The remaining fragments were cultured in vitro for 6 (D6) or 12 (D12) days, containing only minimum essential medium (MEM) or MEM supplemented with alpha-lipoic acid (50, 100, or 250 ng/ml), on an extracellular matrix of agarose gel, in an oven at 38.5ºC. Every 2 days, 100% of the culture medium was replaced. Supplementation with 100 ng/ml was effective for maintaining follicular integrity after 6 days of culture (primordial: 51.28%; development: 36.88%; P0.0001). There was no difference (P0.05) between treatments compared with the non-cultivated control treatment (D0), using the NBT and TBARS assays. Therefore, supplementation of the in vitro culture medium of bovine preantral ovarian follicles with a concentration of 100 ng/ml of alpha-lipoic acid at 6 days of culture was effective for maintaining follicular integrity and, after 6 days, maintaining stable levels of reactive oxygen species.

We evaluated the effect of quercetin on the in vitro culture of bovine ovarian fragments in relation to morphology, development, and oxidative stress. Ovaries (n = 12) from Nelore heifers (n = 6) were used. Each pair of ovaries was divided into nine fragments, and one fragment from each animal was fixed in Bouin solution for 24 h (histology control) or frozen (- 80°C; control for oxidative stress). Other ovarian fragments (n = 8) were distributed into concentrations of 0, 10, 25, and 50 μg/mL of quercetin added to the culture medium for 5 or 10 d. Data were analyzed by chi-square test or ANOVA followed by Tukey's test (P 0.05). Treatment with 25 μg/mL quercetin resulted in the highest proportion of total intact follicles for 5 (67.3%) and 10 d (57.1%); the concentration of 25 μg/mL also presented the best proportion of developing follicles for 5 d (68.7%) and 10 d (62.8%). Treatment with 25 μg/mL quercetin resulted in significant ferric reduction for 10 d of culture, but not for 5 d. No difference (P 0.1) was observed in the production of reactive oxygen species or in the oxidative degradation of lipids between treatments and non-cultivated controls. Treatment with 25 μg/mL quercetin preserved the morphological integrity of the developing follicles for 5 and 10 d of culture, in addition to promoting the best antioxidant potential after 10 d of culture in bovine ovarian fragments.

The goal of this study was to determine the distribution of pre-antral follicles in the ovarian parenchyma of mares. For Experiment 1, each ovary was cut longitudinally at the greater curvature, performing two hemiovaries. After that, six fragments from each hemiovary were obtained, resulting in 12 fragments, which were divided into the innermost region of the parenchyma, the middle region and the outermost region. All the three obtained sections were cut transversally to obtain two fragments from each one. For Experiment 2, each ovary also submitted to a longitudinal cut on the greater curvature, forming two hemiovaries. Each hemiovary was sectioned into four symmetrical fragments, resulting in eight fragments per ovary. The fragments were related as being near to or far from the ovulatory fossa. The fragments of both experiments were immediately fixed in Carnoy for 12 hr and kept in 70% ethanol for 24 hr. Follicles were classified according to the stages of development and for morphological integrity according to oocyte morphology and granulosa cells. After the histological assessment, a total of 1,130 follicles were visualized from Experiment 1, being 1,054 (93.3%) primordial follicles and 76 (4.7%) follicles in development. The innermost region had the highest percentage of pre-antral follicles compared to the other regions (p < .05). The middle and outermost regions showed higher percentages of intact primordial and developing follicles than the innermost region (p < .05). Considering Experiment 2, 938 follicles were found, being 894 (95.3%) primordial and 44 (4.7%) follicles in development. The region near the ovulatory fossa presented higher (58.7%; 551 of 938) follicular concentration compared to the region far from the ovulatory fossa (41.3%; 387 of 938; p < .05). As a conclusion, distribution of pre-antral follicles in the equine ovary has a specific pattern through the parenchyma. Also, the follicular integrity differed in the studied ovarian areas.

Contents This study investigated the effects of different concentrations of FSH (10, 50, 100 and 200 ng/ml) in supplemented MEM+ on the development of equine pre-antral follicles that were cultured in vitro for 2 or 6 days. The ovaries (n = 5) from mares in seasonal anoestrus were collected from a local abattoir. Ten ovarian tissue fragments of approximately 3 × 3 × 1 mm were obtained from each animal. The fragments were cultured in situ for 2 days (D2) or 6 days (D6) in MEM+ or MEM+ supplemented with FSH at four different concentrations, establishing the following 11 groups: control (D0); MEM + (D2); MEM + (D6); MEM + 10 ng/ml of FSH (D2); MEM + 10 ng/ml of FSH (D6); MEM + 50 ng/ml of FSH (D2); MEM + 50 ng/ml of FSH (D6); MEM + 100 ng/ml of FSH (D2); MEM + 100 ng/ml of FSH (D6); MEM + 200 ng/ml of FSH (D2); and MEM + 200 ng/ml of FSH (D6). Follicles were observed in only 9.65% (388 of 4,018) of the histological sections. Of the 861 follicles evaluated, 488 were in the primordial stage, and 373 were in various developmental stages; 59.7% were morphologically normal. Regarding the integrity of the pre-antral follicles, the groups with 100 ng/ml FSH of 2-days culture as well as 50, 100 and 200 ng/ml FSH of 6-days culture provided the best results. In conclusion, the in vitro culture of abattoir-derived equine ovarian fragments presented better morphological integrity when supplemented with FSH for 6 days, in comparison with the MEM culture group. However, no clear effects were observed with FSH regarding the promotion of activation from a primordial to a developing follicle.

O objetivo desse estudo foi determinar a distribuição dos folículos pré-antrais equinos no tecido ovariano, avaliando as regiões interna, média e externa do ovário, bem como as regiões próximas e distantes da fossa ovulatória. Ovários (n=7) de éguas provenientes de abatedouro foram dissecados e seccionados no plano sagital (curvatura maior) resultando em dois hemiovários. Estes foram fragmentados e imersos no fixador Carnoy por 12 horas e mantidos em álcool 70 % até o processamento histológico. No experimento 1 (n=3), delimitou-se à distribuição folicular em regiões ovariana interna (C1), média (C2) e externa (C3). No experimento 2 (n=4) investigou-se a delimitação das regiões próximas (PFO) e distantes (DFO) da fossa ovulatória. A análise estatística para comparação dos dados foi realizada utilizando o teste Exato de Fisher, com nível de 5% de significância através do programa estatístico Minitab® 16.1.1. Encontramos, no primeiro experimento 1.130 folículos, destes 1.054 (93,3 %) eram folículos primordiais e 76 (4,7 %) em desenvolvimento. No experimento 2 foram encontrados 938 folículos, 894 (95,30%) folículos primordiais e 44 (4,7 %) em desenvolvimento. Houve diferença (p

Due to the current understanding of infertility and absence of ovarian activity in the female mule, scientificmaterial regarding their reproductive tract is lacking. Information regarding evaluation of the reproductivesystem, as well as diagnoses of ovarian neoplasms, is scarce for female mules. The purpose of this paperis to report the first case of granulosa cell tumor (GCT) in a female mule, characterized through histology,which suggests evidence of cyclicity. During the breeding season, within the Southern hemisphere, mules’ovaries were obtained from slaughterhouses. Notably, a pair ovarian rated was remarkable in size (16.5cm in diameter), weight (1.92 kg) as well as number of antral follicles. On sagittal section, the tissue wastrabecular and fluid filled. Histology revealed irregular cyst formations full of amphophilic fluid ranging incolor from pinkish to purple. The granulosa cells showed up in abnormal histology, featuring a granulosacell tumor. The contralateral ovary was evaluated, demonstrating anatomy, weight and granulosa cellsfrom normal. Therefore, we have described the first granulosa cells tumor report in a mule via gross andhistological features, which suggests evidence of cyclicity in female mules.

A teratoma is a tumor characterized by a change in the size of the ovary and by presence of well differentiated tissues that are not usual to ovarian stroma. This entity is derived from totipotent cells and early ovarian germ cell differentiation into various tissues. This report aims to describe the morphologic and histologic features of a teratoma found within an antral follicle in the ovary of a mare after slaughter. The ovary was 8 cm in diameter and had a smooth, nodular surface. One antral follicle of 4 cm in diameter containing fluid and lumps of material, both yellow in color, and adhering to a skin, cartilage and bone fragment 1 cm in diameter was visually identified to be attached to the ovarian tissue. Healthy follicles smaller than 3 cm in diameter and hemorrhagic follicle were also observed. Histologic showed normal ovarian tissue containing antral follicles, hemorrhagic follicles and neoformation of multiple tissues. The presence of areas containing epidermis, dermis and skin appendages (sebaceous glands, sweat glands and hair follicles) surrounding cystic cavities filled with keratin, fragments of the cholesterol crystal, and macrophage-containing, waxy sebaceous material was also noted. This paper reports the presence of a teratoma within an equine antral follicle and suggests functionality of the ovary, even in the presence of the teratoma, due to the presence ofactive antral follicles and recent ovulation.