Title page for ETD etd-06132007-113852


Type of Document Dissertation
Author Eljarah, Abdulhakeem Hashim
Author's Email Address aaljar1@lsu.edu
URN etd-06132007-113852
Title Effects of Cryopreservation and Constituents of Semen Extenders on Mitochondrial Function of Bull Spermatozoa
Degree Doctor of Philosophy (Ph.D.)
Department Animal Science (Animal, Dairy, & Poultry Sciences)
Advisory Committee
Advisor Name Title
John Chandler Committee Chair
Cathleen Williams Committee Co-Chair
Dale Paccamonti Committee Member
Fakhri Albagdadi Committee Member
Jill Jenkins Committee Member
John Lynn Committee Member
Huangen Ding Dean's Representative
Keywords
  • Spermatzoa
  • Bovine
  • Cryopreservation
  • Extender
  • Mitochondrial function
Date of Defense 2007-05-15
Availability unrestricted
Abstract
This study investigated the effects of semen extender constituents and cryopreservation on bovine spermatozoal mitochondrial function. Three yearling Holstein bulls were used. Two ejaculates per bull were collected and pooled on a weekly basis for five weeks and extended in four treatments: 1) sodium citrate egg yolk extender with antibiotics (lincomycin, spectinomycin, gentamicin and tylosin); 2) 1 with glycerol; 3) 2 without antibiotics; and 4) 1 without antibiotics. Each was divided into portions for analyses before freezing and after cryopreservation. The pre-freeze and thawed semen were transferred to a 37C water bath, the same assays were performed. In experiment 1, resazurin reduction (RD) was measured spectrophotometrically at sequential 25 minute intervals for 125 minutes. In experiment 2, specific activities of cytochrome c oxidase (CytoCox) and citrate synthase (CS) were measured spectrophotometrically immediately post-thaw and after 125 minutes of incubation. In experiment 3, ATP was measured using luciferin-luciferase assay simultaneously with RD. Total and progressive motilities (TM and PM), progressive (PV), curvilinear (VCL) and pathway (VAP) velocities were measured simultaneously with RD, ATP content and CytoCox and CS using computer assisted semen analysis system (CASA). In experiment 4, the NADH dehydrogenase (ND1) gene of mtDNA was sequenced before and after cryopreservation using PCR. Data were analyzed by least square methods; mean differences were delineated by Tukeys test.

In experiment 1, RD differed among treatments (P<0.05). Cryopreservation decreased (P<0.05) RD, TM, PM, PV, VAP and VCL. Resazurin reduction correlated with PM (r=0.45, P<0.05) and TM (r=0.2, P<0.05). In experiment 2, incubation time and incubation with Triton X100 were sources of variation in CytoCox and CS specific activities (P>0.05). Only CS from spermatozoa incubated with Triton X100 correlated with RD (r=0.22, P<0.05). CytoCox and CS did not correlate with motility parameters. In experiment 3, spermatozoal ATP was not different (P>0.05) among treatments. However, cryopreservation decreased (P<0.05) ATP. Spermatozoal ATP correlated with motility parameters (r0.65) and RD (r=-0.30) (P<0.05). In experiment 4, the frequency of amino acid change was higher (P<0.05) post-thaw in the treatment containing only antibiotics. Cryopreservation, more than extender constituents impacted mitochondrial function of bovine spermatozoa.

Files
  Filename       Size       Approximate Download Time (Hours:Minutes:Seconds) 
 
 28.8 Modem   56K Modem   ISDN (64 Kb)   ISDN (128 Kb)   Higher-speed Access 
  eljarah-thesis.pdf 610.26 Kb 00:02:49 00:01:27 00:01:16 00:00:38 00:00:03

Browse All Available ETDs by ( Author | Department )

If you have questions or technical problems, please Contact LSU-ETD Support.