Please use this identifier to cite or link to this item: http://krishikosh.egranth.ac.in/handle/1/5810082847
Authors: Rajni Kumari
Advisor: Trilok Nanda
Title: Studies on enrichment of X chromosome bearing spermatozoa in buffalo bull semen
Publisher: LUVAS
Language: en
Type: Thesis
Agrotags: null
Keywords: Enrichment, X chromosome bearing spermatozoa, Buffalo bull semen, Density gradient centrifugation,Viability
Abstract: Sex pre-selection has a significant economic impact in livestock industry. Density gradient centrifugation might be an approach to sexing spermatozoa because of the additional DNA content and volume of X chromosome bearing spermatozoa head. The objective of present study was to develop an appropriate density gradient model for enriching X chromosome bearing spermatozoa population in semen of buffalo bull. Density gradient centrifugation (DGC) technique was employed for enriching X chromosome spermatozoa in buffalo bull semen. DGC was carried out using 4 gradient medias viz., Percoll, Optiprep, Ficoll and Sucrose. Pooled semen samples of Murrah buffalo bull were subjected to a total of 24 DGC models (six models per gradient media- based on the density range , number of layers and speed of centrifugation(g) used), swim-up (SU) technique and simple washing in five replicates. The percentage enrichment of X chromosome bearing spermatozoa in all the DGC models was determined by SYBR green based Real Time PCR. Statistical analysis revealed that number of layers and centrifugation speed (g) factors in a density gradient centrifugation model, have significant effect on the percentage enrichment of X chromosome content in semen samples.Three layers, four layers and five layers density gradient centrifugation models showed significant differences (P<0.05) in the sex ratio towards X spermatozoa population in semen (61.72%±0.81, 64.55%±0.26 and 67.31±0.33%) respectively. Density gradient centrifugation models with centrifugation speed (g) 200x g and 300x g also showed significant differences (P<0.05) in the sex ratio towards X spermatozoa population in semen (66.36%±0.25 and 62.69±0.22%) respectively. Based on the results of percentage X enrichment by 24 different density gradient media models, Optiprep DGC model with five layers and centrifugation speed of 200xg was found to yield maximum enrichment (72.4%±1.38). Present study demonstrates that, Optiprep density gradient model with 5 layers and centrifugation at 200xg can be considered as a promising model for enrichment of X spermatozoa from buffalo bull semen based on percentage X enrichment and viability characteristics.
Subject: Animal Biotechnology
Theme: Studies on enrichment of X chromosome bearing spermatozoa in buffalo bull semen
These Type: Ph.D
Issue Date: 2018
Appears in Collections:Thesis

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