The effect of sperm preparation and co-incubation time on in vitro fertilization of bos indicus oocytes

Dode, M. A. N.; Rodovalho, Norma Cléa Mesquita; Ueno, Vanessa Gomes; Fernandes, Carlos Eurico dos Santos

doi:10.1016/s0378-4320(01)00148-8

Cited by 25 publications

(23 citation statements)

References 29 publications

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“…In fact, AR status has been previously used as predictor of both freezability (Molnarova et al, 2006) and in vivo fertility (Birck et al, 2010;Collin et al, 2000;Giritharan et al, 2005) in cattle. However, it has been widely demonstrated that in vitro AR rates depend on both the incubation time (Dode et al, 2002;Fukui et al, 1990;Gliedt et al, 1996;Pavlok et al, 2001) and the culture medium (Januskauskas et al, 2000;Sharma et al, 1999;Vadnais et al, 2005). Our results are consistent with these previous observations, showing a clear interaction between culture medium and incubation time.…”

Section: Discussionsupporting

confidence: 93%

In vitro induction of the acrosome reaction in spermatozoa from endangered Spanish bulls: Effect of breed, culture media and incubation time

et al. 2012

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Section: Discussionsupporting

confidence: 93%

In vitro induction of the acrosome reaction in spermatozoa from endangered Spanish bulls: Effect of breed, culture media and incubation time

et al. 2012

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“…The average number of embryos reaching the blastocyst stage was not influenced by treatment (Table 3) and was comparable to those reported in the literature (Dode et al, 2002;Chaubal et al, 2007;Sales, 2010). Since the number of oocytes derived from the Holstein heifers was low, it was not possible to evaluate the main effect of breed on the final in vitro embryo development.…”

Section: Resultssupporting

confidence: 88%

Association of recombinant bovine somatotropin (rBST) with equine chorionic gonadotropin (eCG) on antral follicle count and oocyte production in Holstein and Tabapuã heifers

et al. 2012

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-The objective of this study was to investigate whether the use of rbST and eCG prior to ultrasound-guided follicular aspiration (OPU) improves oocyte yield and quality in Tabapuã and Holstein heifers. The study was conducted in two phases, 20 days apart, in a change-over design. The dominant follicle was ablated two days (D-2) before two treatments: stimulation (6 Holstein and 8 Tabapuã), 500 mg of rbST (Boostin ® ) on D0 and 500 IU of eCG (Folligon) on D2; and control (6 Holstein and 8 Tabapuã), in which heifers received injections of the excipient. Heifers were aspirated on D4. Oocytes were subjected to a well established commercial in vitro embryo production protocol (Vitrogen ® ) and embryos were evaluated seven days after fertilization. There was an effect from the interaction of treatment and breed, so that hormonal stimulation increased antral follicle count (2-8 mm) in Tabapuã (29.9±2.6 to 41.4±2.6), but not in Holstein heifers (14.4±2.6 to 15.5±2.6). Tabapuã heifers had higher mean antral follicle count than Holsteins (35.6±1.8 vs. 15.0±2.1). The number of viable oocytes was not increased by stimulation in Tabapuã (from 4.7±1.0 to 5.2±1.1 in control and simulation, respectively) or in Holstein heifers (1.3±1.9 to 2.0±1.6 in control and simulation, respectively). There was no difference in the percentage of heifers with more than five viable oocytes in the group treated (33 vs 27%). The number of blastocysts was not affected by treatment (1.75 vs. 1.00 in hormonal stimulation and control, respectively). The increase in antral follicle count in the stimulated Tabapuã heifers did not reflect upon oocyte yield. The differential breed response to the hormonal treatment underscores the need for additional tests, especially for the Holstein breed, in order to enhance OPU efficiency.

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“…These techniques are used for removing seminal plasma, dead cells, abnormal sperm, cryoprotective agents, and other factors [15]. The techniques include Percoll density gradient centrifugation [4,7,12-15,19,22], swim-up migration [4,7,15], washing by centrifugation [7,15], glass wool filtration [15], and many other.…”

Section: Discussionmentioning

confidence: 99%

“…Also, Percoll gradient is the preferred method to prepare frozen-thawed bovine sperm for IVF because it yields higher motility and a higher number of insemination doses [4]. In addition, no differences in fertilization rates have been reported among Percoll gradient and other typical sperm separation methods [7]. …”

Section: Discussionmentioning

confidence: 99%

Transmission electron microscopy for characterization of acrosomal damage after Percoll gradient centrifugation of cryopreserved bovine spermatozoa

et al. 2011

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The objective of this study was to characterize acrosomal ultrastructure following discontinuous Percoll gradient centrifugation of cryopreserved bovine sperm. Semen was collected from six bulls of different breeds and three ejaculates per bull were evaluated. Frozen semen samples were thawed and the acrosomal region of sperm cells was evaluated by transmission electron microscopy (TEM) before (n = 18) and after (n = 18) Percoll centrifugation. The evaluation of 20 sperm heads from each of the 36 samples analyzed ensured that a large number of cells were investigated. The data were subjected to analysis of variance at a level of significance of 5%. Percoll centrifugation reduced the percentage of sperm exhibiting normal acrosomes (from 61.77 to 30.24%), reduced the percentage of sperm presenting atypical acrosome reactions (from 28.38 to 4.84%) and increased the percentage of sperm exhibiting damage in the acrosome (from 6.14 to 64.26%). The percentage of sperm with typical acrosome reactions was not significantly different before (3.70%) and after (0.67%) centrifugation. TEM distinguished four different types of acrosomal status and enabled ultrastructural characterization of acrosomal injuries. The percentage of sperm exhibiting normal acrosomes decreased and damage in the acrosome was the most frequent acrosomal injury with the Percoll gradient centrifugation protocol utilized.

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The effect of sperm preparation and co-incubation time on in vitro fertilization of bos indicus oocytes

Cited by 25 publications

References 29 publications

In vitro induction of the acrosome reaction in spermatozoa from endangered Spanish bulls: Effect of breed, culture media and incubation time

In vitro induction of the acrosome reaction in spermatozoa from endangered Spanish bulls: Effect of breed, culture media and incubation time

Association of recombinant bovine somatotropin (rBST) with equine chorionic gonadotropin (eCG) on antral follicle count and oocyte production in Holstein and Tabapuã heifers

Transmission electron microscopy for characterization of acrosomal damage after Percoll gradient centrifugation of cryopreserved bovine spermatozoa

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