Leakage of Phosphatases and Fertility of Buck Semen Cryopreserved under Different Freezing Modes

  • Sunanda Sharma College of Veterinary & Animal Science, Rajasthan University of Veterinary & Animal Sciences, Bikaner-334001.
  • Narendra Kumar Sharma Mobile Veterinary Surgery & Infertility Unit, Animal Husbandry Department, Bikaner-334001, India
  • Nalini Kataria Department of Veterinary Physiology College of Veterinary & Animal Science, Rajasthan University of Veterinary & Animal Sciences, Bikaner-334001, India.
  • N K Sinha Division of Reproduction, Physiology and Shelter Management, Central Institute for Research on Goats Makhdoom-Farah-Mathura, India.
Keywords: Buck Semen, Freezing rates, Seminal Plasma phosphatases, Semen Cryopreservation, ACP, AKP

Abstract

Present study was conducted on 160 ejaculates collected at weekly interval by artificial vagina method from 13 adult Sirohi bucks. Pooled ejaculates were diluted with Tris-egg yolk-citric acid-fructose-glycerol extender (1:4), filled and sealed in straws. Few straws of diluted semen were thawed (40° C/15 seconds) and assessed for acid and alkaline phosphatases (ACP and AKP) in seminal plasma of diluted semen (Control Group). Remaining semen straws were randomly grouped to constitute freezing mode groups (M1, M2, M2, and M4) and processed further for cryo-preservation of semen. Accordingly diluted semen straws were cooled @-4° C/minute from 25° C up to 5° C thereafter equilibrated for 2 hours and frozen up to -160° C @ 15, 20, 25 and 300 C/minute for M1, M2, M3 and M4 groups respectively. These frozen straws were hold at this temperature for 2 minutes then stored separately in LN2. After 7 days of storage, straws from each freezing mode group were thawed and assessed for ACP and AKP in seminal plasma. In vivo fertility trials were also conducted with straws of control (fresh diluted semen) as well as freezing mode groups (frozen at different freezing rates). Least square analysis of variance for the data obtained revealed highly significant (P ≤ 0.01) rise in the seminal plasma ACP and AKP enzyme levels in frozen thawed semen as compared to that in fresh diluted semen. The Values of ACP and AKP also differed significantly (P ≤ 0.05) among all the freezing mode groups wherein lowest values of ACP were observed in M3 group followed by M4, M2 and M1 groups in increasing order whereas, lowest values of AKP were observed in M3 followed by M2, M4 and M1 groups in increasing order. Highest fertility rates were observed with semen from M3 followed by M2, M4 and M1 groups. On the basis of enzyme leakage and in-vivo fertility trials, the optimum freezing rate for cryopreservation protocol was arrived at 25° C/minute.

DOI: http://dx.doi.org/10.3126/ijasbt.v1i4.8913

 Int J Appl Sci Biotechnol, Vol. 1(4): 202-207

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Author Biographies

Sunanda Sharma, College of Veterinary & Animal Science, Rajasthan University of Veterinary & Animal Sciences, Bikaner-334001.

Graduated in 1990 from College of Veterinary & Animal Science, Bikaner and awarded B.V.Sc.& A.H. degree with 60.92 percent marks by Rajasthan Agricultural University, Bikaner, India,  Dr. Sunanda Sharma began her carrier in 1990 at College of Veterinary & Animal Science, Bikaner as IRPP fellow during her post-graduation research in Veterinary Obstetrics & Gynaecology and submitted research thesis for that she was awarded M.V.Sc. (Animal Reproduction, Obstetrics & Gynaecology) degree  in July 1994 with 86.47 percent marks by Rajasthan Agricultural University, Bikaner, India. She joined and worked in Animal Husbandry Department, Government of Rajasthan wherein for 6 years she imparted training on "Artificial Insemination, Handling and Maintanance of frozen semen and LN2” to Veterinary Officers, Para-Vets and Village Level Inseminators (Gopal) . Afterward she earned vast clinical experience for more than 11 years as Veterinary Officer and Incharge of Government Veterinary Hospital at different places. She did doctoral research in buck semenology and cryo-preservation for 2 years at Central Institute for Research on Goats (ICAR), Makhdoom-Farah-Mathura and submitted thesis for that she was awarded Doctor of Philosophy (Animal Reproduction, Obstetrics & Gynaecology) degree in 2004 with OGPA 4.00 out of 4.00 (90% marks) by Rajasthan Agricultural University, Bikaner, India . She earned more than 20 Year experience as clinician, trainer, teacher, researcher and event organizer in the displine of Veterinary Science and Animal Husbandry. Presently she is working as Faculty Member at Department of Veterinary Obstetrics & gynaecology, College of Veterinary and Animal Science, Rajasthan University of Veterinary & Animal Sciences, Bikaner, India wherein she is engaged in conducting and guiding post-graduate research as University approved guide; teaching theory, practical and clinical classes to graduate, post-graduate and diploma courses at college of Veterinary & Animal Science, Rajasthan University of Veterinary & Animal Sciences, Bikaner, India. She had published 2 research thesis by her name, 10 research/clinical papers in journals of repute, 5 teaching manuals, 8 papers in international and national conferences, 6 technical trifold folders and  One Book. She is also contributing voluntary services as Editor, Editorial board member in 10 journals and Reviewer in 20 International Journals of repute

Narendra Kumar Sharma, Mobile Veterinary Surgery & Infertility Unit, Animal Husbandry Department, Bikaner-334001, India
Senior Veterinary Officer, Incharge-Mobile Veterinary Surgery and Infertilty Unit, Animal Husbandry Department, Bikaner. Graduated (B.V.Sc. & AH) in 1984 with 76% marks, thereafter joined as Veterinary Officer at Civil Veterinary hospital. Master's Degree (MVSc.) in Veterinary Surgery in 1995. Had more than 29 years clinical experience in Veterinary Medicine, Veterinary Surgery, Veterinary Obst. & gynaecology, Animal Reproduction. Reviewer in 6 journals of international repute.
Nalini Kataria, Department of Veterinary Physiology College of Veterinary & Animal Science, Rajasthan University of Veterinary & Animal Sciences, Bikaner-334001, India.

as Faculty member engaged in teaching Veterinary Physiology at college of veterinary & Animal Science, Bikaner.

Editor and reviewer in ELBS Bioflux Journal

N K Sinha, Division of Reproduction, Physiology and Shelter Management, Central Institute for Research on Goats Makhdoom-Farah-Mathura, India.

Former Principal Scientist (Animal Reproduction) at Division of Reproduction, Physiology and Shelter Management,Central Institute for Research on GoatsMakhdoom-Farah-Mathura, India.

Co-Advisor for Ph.D. research work of Dr.Sunanda Sharma (Corresponding author of this paper.) 

Published
2013-12-21
How to Cite
Sharma, S., Sharma, N., Kataria, N., & Sinha, N. (2013). Leakage of Phosphatases and Fertility of Buck Semen Cryopreserved under Different Freezing Modes. International Journal of Applied Sciences and Biotechnology, 1(4), 202-207. https://doi.org/10.3126/ijasbt.v1i4.8913
Section
Research Articles