<div style="text-align: justify;">The aim of the present study was to produce goat embryo in different culture media through in vitro fertilization using cryopreserved black Bengal buck semen. Total 1265 fresh cumulus oocyte complexes (COCs) were collected by aspiration method with a 19 gauge hypodermic needle, washed 5-6 times and cultured in maturation media maintaining 5% CO2 level at 38.5&ordm;C with maximum humidity in a incubator. After 27 h of incubation cumulus cells were stripped off from matured oocytes and transferred to acidified Tyrode&rsquo;s medium for zona thinning and co-incubated with in vitro capacitated sperms for fertilization in Fert-BO media. In the experiment I, fresh buck semen and in experiment II, frozen buck semen was used for in vitro fertilization after in vitro processing. After 5 h of co-incubation, presumptive zygotes were washed and co-incubated with oviductal cells in three different culture media (RVCL, mSOF, KSOM) for further development. In fresh group cleavage (%) were 37.76 &plusmn; 2.98, 39.60 &plusmn; 1.75, 29.01 &plusmn; 1.74, and morula formation (%) were 7.72 &plusmn; 3.38, 6.03 &plusmn; 1.29, and 3.00 &plusmn; 3.00 in RVCL, mSOF and KSOM media respectively. However, in frozen group cleavage (%) were 29.17 &plusmn; 2.56, 27.70 &plusmn; 2.31, 24.17 &plusmn; 1.44 in RVCL, mSOF and KSOM media respectively and morula formation (%) was 2.93 &plusmn; 0.97 only in RVCL media. These results indicate that cryopreserved black Bengal buck semen have competence to produce embryos and could be used for embryo development in RVCL media through in vitro fertilization.&nbsp;</div>

<div style="text-align: justify;">In vitro Development of Preimplantation Caprine Embryo using Cryopreserved Black Bengal Buck Semen</div>

Background: Production of female offspring is vital for maximizing milk yield in the dairy industry, making embryo sexing essential. Metabolic differences between male and female embryos may be reflected in the culture medium (CM). This study explored non-invasive sex detection of in vitro produced buffalo embryos by analyzing metabolites in spent CM. 
 
Methods: Buffalo ovaries were collected from a slaughterhouse and oocytes were matured and fertilized in vitro. Presumptive zygotes were cultured in either a sequential medium, Synthetic oviduct fluid (SOF) or a single-step medium, Potassium Simplex Optimized Medium (KSOM). After 5 days, morulae were placed in 25 &micro;l of CM. On day 7, embryos and spent medium were collected and stored at -20oC. Embryo sex was determined using PCR. Spent media were analyzed using Attenuated Total Reflection - Fourier Transform Infrared Spectroscopy (ATR-FTIR) spectroscopy to detect sex-specific metabolites. 
 
Result: This approach enabled non-invasive identification of embryo sex and revealed the impact of different culture media on sex ratio. The findings highlight the potential of spent CM analysis for predicting embryo sex, offering significant benefits for the dairy industry.

Comparative Analysis of Culture Media and ATR-FTIR Metabolomics for Non-invasive Sex Determination in In vitro Produced Buffalo Embryos