Relationship between the concentration of anti-Müllerian hormone and antral follicle count in pasture-managed Holstein cows in the high tropics.

We evaluated the relationship between plasma levels of anti-Müllerian hormone (AMH) and the number of antral follicles at the restart of the follicular wave in crossbred Holstein cows reared under extensive grazing systems over 2500 m above sea level. The study included 140 cows from 15 farms that were in average at the 75.3 ± 2.10 d post partum. Animals were synchronized according to the following regime: day 0 = intravaginal progesterone releasing device (IPD) + estradiol benzoate (EB); day 7: withdrawal of IPD + prostaglandin; and day 8: EB, for restart of the follicular wave on day 11. On this day 11, antral follicle counts (AFCs) were made by transrectal ultrasound, and a plasma sample was taken for the determination of AMH. The mean AMH plasma level was 0.06 ± 0.03 ng/ml and the mean AFC was 17.26 ± 0.38 follicles. A strong positive linear correlation was found between these two variables (r = 0.783, r = 0.613, P < 0.0001). Cows were categorized according to AMH concentration as high (>0.09 ng/ml), intermediate (0.09-0.05 ng/ml) or low (<0.05 ng/ml). Cows with high AMH presented a higher AFC (25.0 ± 2.21 follicles) than those with low AMH (14.08 ± 2.68 follicles; P < 0.001. Our results suggest that the cut-off value of AMH = 0.09 ng/ml may be useful for selecting donors in multiple ovulation embryo transfer programs involving cows with these characteristics. Our data further suggest that AMH plasma concentration correlates with AFC and can be used as an endocrine biomarker of the number of antral follicles present at a given moment of the estrous cycle in crossbred Holstein cows raised at altitudes above 2500 m.

Lunar Cycle Influences Reproductive Performance of Crossbred Brahman Cows Under Tropical Conditions.

Evidence has accumulated over the years indicating that the moon influences some aspects of the reproductive activity in animals and humans. However, little is known about the influence of the lunar cycle on the reproductive performance of cows under tropical conditions, where the environment strongly affects reproduction. This retrospective study was conducted with the aim of assessing the influence of the lunar cycle on some reproductive traits of tropical crossbred cows managed in a pasture-based system. Data from 5869 reproductive records from two commercial farms localized in the Maracaibo Lake Basin of Zulia State, Venezuela, were analyzed. Variables studied were first service conception rate, calving frequency, first postpartum estrous frequency, and pregnancy frequency. In addition to the lunar cycle, the effects of farm, season, and predominant breed were also considered. Data were analyzed using logistic regression and general linear model from SAS. First service conception was affected by lunar phases and predominant breed, but not by farm or season. For frequencies of calving, first postpartum estrus, and pregnancy, there was no main effect of farm, season, and predominant breed, whereas the effect of lunar phases was highly significant. First service conception was significantly greater in waning than in crescent phase of the lunar cycle. Frequencies of calving, first estrus, and pregnancy were highly correlated and showed greater figures around full moon and new moon. In conclusion, lunar cycle influenced first service conception, attaining greater values in the waning phase of the moon cycle. Frequencies of calving, first postpartum estrus, and pregnancy in crossbred cows showed a clear bimodal rhythm, whose greatest values coincided with new moon and full moon.

Intraovarian influence of bovine corpus luteum on oocyte morphometry and developmental competence, embryo production and cryotolerance.

Three experiments were conducted to determine influence of the bovine corpus luteum (CL) on morphometric and functional characteristics of oocytes, and subsequent embryonic development. Cumulus-oocyte complexes were aspirated from two types of cows: 1) with a CL in one ovary (CL+) and without a CL in the contralateral ovary (CL-), 2) and from cows without CL in either ovary (C). Intracellular activity of the enzyme glucose-6-phosphate dehydrogenase (G6PDH), oocyte diameter and thickness of the zona pellucida were determined (Experiment 1). Then, the rate of in vitro oocyte maturation for each ovarian category was evaluated and oocyte diameter and zona pellucida thickness were measured after maturation (Experiment 2). In Experiment 3, in vitro embryo production and cryotolerance were assessed. The oocyte diameter was greater (P < 0.01) and the zona pellucida was thinner in CL+ than in CL- (P > 0.05) or C (P = 0.0131) ovaries. Activity of G6PDH was lower in oocytes from CL+ than CL- (P < 0.01) and C (P = 0.0148) ovaries. Rate of oocyte maturation, oocyte diameter and thickness of the zona pellucida after maturation did not differ among groups. Rate of cleavage was greater in zygotes from CL+ than from CL- or C (P < 0.01); and CL+ ovaries produced more total embryos on day 7 (P < 0.05) and more blastocysts (P < 0.01) than CL- and C ovaries. Rate of expansion and hatching of day-7 vitrified-warmed blastocysts at 24 and 48 h of culture did not differ among groups. In conclusion, oocytes collected from CL+ ovaries were larger and metabolically more prepared to continue maturation than those from ovaries lacking a CL. Also, rates of cleavage and yield of blastocysts were greater for oocytes from CL+ ovaries than from CL- and C ovaries. These findings indicate that a CL influenced oocyte developmental competence and embryonic development, presumably through intraovarian interactions.

Evaluación de la capacidad de desarrollo in vitro de ovocitos bovinos provenientes de vacas con predominancia fenotípica Bos taurus y Bos indicus / Evaluation of in vitro development capacity of bovine oocytes obtained from predominantly Bos taurus and Bos indicus cows

El objetivo de este estudio fue evaluar la capacidad de desarrollo in vitro de ovocitos bovinos de vacas mestizas B. taurus y B. indicus. Los ovocitos fueron recuperados de ovarios de hembras bovinas provenientes de un matadero comercial. Para la obtención de los complejos cumulus-ovocitos (CCO) se realizó la técnica de slicing, seleccionando los ovocitos que tenían al menos una capa de células del cumulus y un citoplasma homogéneo. Los ovocitos seleccionados fueron madurados y fecundados in vitro (MIV-FIV). Se utilizó semen de un toro Brahman puro (B. indicus). Para la evaluación de la MIV y FIV todos los ovocitos se fijaron por al menos 24 h a 4°C en solución metanol-ácido acético (3:1) y teñidos con aceto-orceína al 1,1%. La tasa de maduración de ovocitos de vacas con predominancia fenotípica B. indicus fue del 66,17% mientras que las vacas con predominancia fenotípica B. taurus alcanzaron un 50,94% (P>0,05). En cuanto a la tasa de fecundación se obtuvo un 14,28 y 35,72% de ovocitos penetrados normalmente y anormales, respectivamente, para el grupo de ovocitos con predominancia fenotípica B. indicus. Mientras que para vacas con predominancia fenotípica B. taurus, un 10,22% correspondió a ovocitos penetrados normales y 19,31% de ovocitos penetrados anormales, sin encontrar diferencias estadísticamente significativas en ambos grupos. Los presentes resultados, tanto para la progresión meiótica como para las tasas de fecundación, indican que los ovocitos de vacas mestizas con predominancia fenotípica B. indicus son más competentes en las primeras etapas de desarrollo in vitro que los ovocitos de vacas mestizas con predominancia fenotípica B. taurus.

The aim of this study was to evaluate the in vitro development capacity of bovine oocytes from crossbred B. taurus and B. indicus cows. Oocytes from bovine cows were collected from commercial slaughterhouse. The cumulus-oocyte complex (COC) ovaries were obtained by Slicing technique, selecting those oocytes that had 2 to 3 layers of cumulus cells and homogeneous cytoplasm. After selection oocytes proceed with maturation (IVM) and fertilization in vitro (IVF). It used semen from a pure Brahman bull (B. indicus). For the assessment of IVM as for IVF oocytes were fixed for about 24 hours at 4°C in methanol-acetic acid (3:1) solution and stained with 1.1% aceto-orcein. The maturation rate of oocytes from cows with B. indicus phenotypic predominance was 66.17%, whereas cows with B. taurus phenotypic predominance 50.94% (P>0.05). Fertilization rate obtained in B. indicus phenotypic predominance group was 14.28% of oocytes normal penetrated and abnormal penetrated 35.72%, for cows with a phenotypic predominance B. taurus oocytes normal penetrated were 10.22% and 19.31% of abnormal oocytes penetrated. In conclusion, the present results indicate that oocytes from cows with phenotypic predominance B. indicus are more competent in the early stages of development in vitro than oocytes from cows with phenotypic predominance B. taurus.

Time course of follicular development after bovine ovarian tissue transplantation in male non-obese diabetic severe combined immunodeficient mice.

OBJECTIVE: The aim of this study was to determine, with a bovine model, the appropriate interval for xenografted adult and newborn ovarian tissue to develop gonadotropin-responsive follicles. DESIGN: Controlled experiment. SETTING: Academic research laboratory. ANIMAL(S): Male non-obese diabetic (NOD) severe combined immunodeficient (SCID) mice (n = 20) were hosts of bovine ovarian tissue. Two dairy calves and one adult beef cow were donors of ovarian tissue. INTERVENTION(S): Newborn and adult bovine ovarian cortical pieces were transplanted to the SC space of intact male NOD SCID mice. Grafts were recovered after euthanasia at intervals after transplantation. MAIN OUTCOME MEASURE(S): Microscopic examination of histologic sections to determine proportions of growing follicles. RESULT(S): There was an increase in the proportion of primary and secondary follicles on day 55 after surgery for the cow and on day 124 after surgery for calf tissue compared with nongrafted and xenografted ovarian tissues recovered at previous intervals. These observed increases were accompanied by decreases in proportions of primordial follicles. Results suggest a sudden increase in the proportion of primary and secondary follicles due to progressive development of primordial follicles. CONCLUSION(S): In the NOD SCID mouse, bovine follicles survived xenotransplantation and underwent development. A longer interval was required for ovarian follicular development in calf tissues compared with that in adult cow ovarian tissues after xenotransplantation.

Development of antral follicles in cryopreserved cat ovarian tissue transplanted to immunodeficient mice.

Ovarian cortex cryopreservation and xenotransplantation into immunodeficient mice represents a potential means for female germplasm conservation and an immediate model for investigation of folliculogenesis. The objectives of this study were to: (1) assess follicle survival after cryopreservation and transplantation of cat ovarian tissue into non-obese diabetic severely combined immunodeficient (NOD SCID) mice; and (2) evaluate the effects of gonadotropin treatments on follicular development in the transplanted tissue. Slices from the cat ovarian cortex were frozen and after thawing, transplanted under each kidney capsule of castrated male NOD SCID mice (eight xenografts in four mice). Sixty-two days after surgery, mice were randomly assigned (two per group) to gonadotropin-treated (eCG and hCG 88 h later) or control (saline-treated) groups. Twenty-four hours after the last injection, ovarian tissue was recovered and processed for histology. Fresh ovarian tissue from the same original source was similarly processed. Follicles were counted, measured, and classified as primordial, primary, secondary, or antral. Immunoreactive proliferating cell nuclear antigen (PCNA) stain was used to assess follicle viability. Microscopic examination revealed no evidence of necrosis or fibrosis. The grafts were well-vascularized, with follicles at all stages of development. Numbers of follicles in the transplanted tissue were markedly reduced compared to fresh tissue, with approximately 10% of follicles surviving freezing and transplantation procedures. Growing follicles positive for PCNA were found in all xenografts. Gonadotropin treatment did not alter the proportion of resting to growing follicles or mean follicle diameter by comparison with controls from untreated mice. By contrast, luteinization, but not ovulation, of antral follicles was observed only in grafts from treated mice. In summary, frozen-thawed cat ovarian cortex tissue not only survived xenotransplantation, it also contained follicles able to grow to antral stages. Exogenous gonadotropin treatment in this model resulted in luteinization of antral follicles but enhancement of follicular growth and ovulation did not occur.