Sex determination of buffalo embryos (<i>Bubalus bubalis</i>) by polymerase chain reaction

Manna, Laura; Neglia, Gianluca; Marino, Marcella; Palo, R. Di; Zicarelli, L.

doi:10.1017/s0967199403001035

Cited by 17 publications

(11 citation statements)

References 29 publications

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“…After digestion, proteinase K was inactivated at 958C for 10 min. Two sets of PCR primers were used to determine embryo sex: Y-chromosome specific primers (BRY1a), and bovine specific satellite sequence primers (Sat1) [15]. The PCR reactions were conducted in a total volume of 25 ll containing 8 ll of the proteinase K digested sample, 13 Gotaq Flexi buffer, 1 IU of Gotaq (Promega, Madison, WI), 1.25 mM MgCl 2 , 0.1 mM dNTP, 1 ng/ll BRY primers, and 0.2 ng/ll Sat primers.…”

Section: Embryo Sexing By Pcrmentioning

confidence: 99%

Can Bovine In Vitro-Matured Oocytes Selectively Process X- or Y-Sorted Sperm Differentially?1

Bermejo-Álvarez¹,

Rizos²,

Rath³

et al. 2008

Biology of Reproduction

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It has been reported that the mammalian female could have a preconceptual influence on the sex of her offspring, and it has been hypothesized that this influence could go some way toward accounting for the reported lower fertility following insemination with sex-sorted sperm. To test whether in vitro matured oocytes are able to select X-or Y-bearing spermatozoa following in vitro fertilization (IVF), we fertilized in vitro 1788 oocytes with Xsorted semen, Y-sorted semen, a mix of X-and Y-sorted semen, and unsorted semen from the same bull, and cultured until Day 9. Fertility was assessed by recording cleavage rate at 48 h postinsemination (hpi) and blastocyst development until Day 9. Embryos were sexed at the two-to four-cell stage and the blastocyst stage. The proportion of zygotes cleaving at 48 hpi was not different between X-and Y-sorted groups and the mix of Xand Y-sorted semen group; however, all were significantly lower than the unsorted group (P , 0.001). Blastocyst yield on Day 6 was significantly higher (P 0.01) in the control group compared with the rest of the groups. Cumulative blastocyst yields on Days 7, 8, and 9 were also significantly higher (P 0.01) in the unsorted group compared with the sorted groups. The proportion of female and male two-to four-cell embryos obtained following IVF with X-and Y-sorted sperm was 88% and 89%, respectively and the sex ratio at the two-to four-cell stage was not different following IVF with unsorted or sorted/recombined sperm (56.9% males vs. 57% males, respectively). At the blastocyst stage, similar percentages were obtained. In conclusion, the differences in cleavage and blastocyst development using sorted versus unsorted sperm are not due to the oocyte preferentially selecting sperm of one sex over another, but are more likely due to spermatic damage caused by the sorting procedure.animal reproduction technology, fertilization, in vitro fertilization, male and female bovine embryos, sex-sorted sperm

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Section: Embryo Sexing By Pcrmentioning

confidence: 99%

Can Bovine In Vitro-Matured Oocytes Selectively Process X- or Y-Sorted Sperm Differentially?1

Bermejo-Álvarez¹,

Rizos²,

Rath³

et al. 2008

Biology of Reproduction

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“…In buffaloes, bovine Y‐chromosome‐specific primers have been used to test in vitro produced buffalo embryos for gender at different stage of development (Rao et al. 1993; Manna et al. 2003).…”

Section: Female Reproductive Function and Assisted Reproductive Technmentioning

confidence: 99%

Reproduction in the Water Buffalo

Presicce

2007

Reprod Domestic Animals

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In this paper, an account of various aspects related to buffalo reproduction are given. Fundamental concepts of the reproductive physiology as well as manipulation of the reproductive function will be presented. This will include an overview of the most recent developments of the oestrous cycle and the ovulation control, new strategies of reproductive management for the improvement of genetic gain and the application of newly developed reproductive technologies, such as in vitro embryo production, embryo and sperm sexing and cloning.

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“…The most common approach in sexing embryos involves the co-amplification of the Y chromosomespecific sequence, which contains the Y-linked genes (SRY) and an autosomal sequence that acts as a control for the presence of DNA (Peura et al, 1991;Appa Rao et al, 1993;Macháty et al, 1993;Park et al, 2001;Manna et al, 2003).…”

Section: Discussionmentioning

confidence: 99%

Sex determining of cat embryo and some feline species

Ciani

Cocchia

Rizzo

et al. 2008

Zygote

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SummarySex identification in mammalian preimplantation embryos is a technique that is used currently for development of the embryo transfer industry for zootechnical animals and is, therefore, a resource for biodiversity preservation. The aim of the present study was to establish a rapid and reliable method for the sexing of preimplantation embryos in domestic cats. Here we describe the use of nested PCR identify Y chromosome-linked markers when starting from small amounts of DNA and test the method for the purpose of sexing different species of wild felids. To evaluate the efficiency of the primers, PCR analysis were performed first in blood samples of sex-known domestic cats. Cat embryos were produced both in vitro and in vivo and the blastocysts were biopsied. A Magnetic Resin System was used to capture a consistent amount of DNA from embryo biopsy and wild felid hairs. The results from nested PCR applied on cat blood that corresponded to the phenotypical sex. Nested PCR was also applied to 37 embryo biopsies and the final result was: 21 males and 16 females. Furthermore, β-actin was amplified in each sample, as a positive control for DNA presence. Subsequently, nested PCR was performed on blood and hair samples from some wild felines and again the genotyping results and phenotype sex corresponded. The data show that this method is a rapid and repeatable option for sex determination in domestic cat embryos and some wild felids and that a small amount of cells is sufficient to obtain a reliable result. This technique, therefore, affords investigators a new approach that they can insert in the safeguard programmes of felida biodiversity.

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Sex determination of buffalo embryos (Bubalus bubalis) by polymerase chain reaction

Cited by 17 publications

References 29 publications

Can Bovine In Vitro-Matured Oocytes Selectively Process X- or Y-Sorted Sperm Differentially?1

Can Bovine In Vitro-Matured Oocytes Selectively Process X- or Y-Sorted Sperm Differentially?1

Reproduction in the Water Buffalo

Sex determining of cat embryo and some feline species

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