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In Vitro Culture (ivc) of Buffalo Embryos

Dr. M.A. Yahia Khandaker
Professor
Department of Animal Breeding and Genetics, BAU, Mymensingh-2202

N. Jahan
Department of Animal Breeding and Genetics, BAU, Mymensingh-2202

M. T. Reza
Department of Animal Breeding and Genetics, BAU, Mymensingh-2202

A. N. M I. Rahman
Department of Animal Breeding and Genetics, BAU, Mymensingh-2202

Abstract/ Executive Summary:

The present research has been undertaken to explore the collection and evaluation of slaughter house ovaries, follicles and COCs, maturation of buffalo oocytes by using bovine follicular fluid (BFF) and bovine serum (BSA) as well as subsequent fertilization using buffalo frozen semen. The results obtained from the experiment shown that there were no significant (p<0.05) difference in the parameters between left and right ovaries except length. The length (cm) was distinctly higher in case of right ovaries (2.32 ± 0.06) compared to left ovaries (2.14 ± 0.085). Normal quality COCs (grade A and grade B) were found significantly (p<0.01) higher in type-1 than those of type-II. Total number of observed follicles aspirated follicles, total number of cumulus-oocyte-complexes (COCs) and number of normal COCs were significantly (p<0.05) decreased when corpus luteum present. Cumulus oocyte complexes (COCs) were collected by aspiration of slaughterhouse buffalo ovaries. After maturation in TCM 199 supplemented with one of 10% BFF, 5% BSA or without supplementation (control), the COCs were fertilized in Brackett and Oliphant (BO) medium using capacitated frozen buffalo spermatozoa. It was observed that the percentage of COCs reached to M-II stages were 40.78 ± 3.84, 65.74 ± 2.39 and 67.52 ± 0.85; normal fertilization (formation of 2 pronuclei) were 23.28 ± 3.00, 29.30 ± 0.73 and 30.52 ± 1.21 for control 10% BFF and 5% BSA supplementation, respectively. The results indicated that the rate of maturation could be significantly increased (p<0.05) by using 10% BFF or 5% BSA to that of control but the supplements showed similar results (p<0.05). In fertilization, it was observed that buffalo frozen spermatozoa could fertilize the buffalo oocytes and the fertilization rate was 23.28% to 30.52%. The fertilizer rates were significantly increased (p<0.05) when 10% BFF or 5% BSA was supplemented in maturation media. Therefore, it could be concluded that buffalo oocytes might be fertilized by using capacitated buffalo spermatozoa and both 10% BFf and 5% BSA could be supplemented in maturation media to enhance the maturation rate as fertilization of buffalo oocytes and the results suggest that the culture condition for IVC was found optimum and the grade A COCs might be suitable for buffalo embryo production.

Keywords: Buffalo, Embryo, oocytes, Artificial insemination, IVP

Location: Department of Animal breeding and genetics

Start Date: 01-07-2010

End Date: 30-06-2013

Program Area: Variety and Species

Commodity: Buffalo

Objectives:

1) To obtain the efficient culture condition for in vitro fertilization (IVF) of Buffalo octets 2) To establish the suitable culture condition for in vitro culture (IVC) of buffalo embryos 3) To test the viability of the in vitro developed blastocysts 4) To build up knowledge base in the in vitro production (IVP) of buffalo embryos

Methodology:

Methodology was followed the steps: 1) Experimental site selection 2) Collection and processing of ovaries 3) Collection and grading of cumulus-oocyte-complexes (COCs) 4) In vitro maturation (IVM) of oocytes 5) In vitro fertilization (IVF) 6) Checking the fertilization rate

Source of Information: Bangladesh Agricultural University Research Progress, Volume-22, 2011. Proceedings of the workshop of BAU Research Progress, 02-03 April 2012 and Annual workshop on BAU research Progress: 2010-2011; BAURES, 02-03 April 2012, BAU, Mymensingh, 02-03-2012

Article Link (Online Journal):

Funding Source:

1.	Budget: 2100000

Total Budget (Tk.) : 2100000

Achievement/Findings:

Buffalo oocytes might be fertilized by using capacitated buffalo spermatozoa and both 10% BFf and 5% BSA could be supplemented in maturation media to enhance the maturation rate as fertilization of buffalo oocytes and the results suggest that the culture condition for IVC was found optimum and the grade A COCs might be suitable for buffalo embryo production.

Knowledge Management: Report/Proceedings