title: Effect of incubation on freezability of cholesterol-loaded cyclodextrin treated buffalo (Bubalus bubalis) spermatozoa.
abstract: OBJECTIVE
  The aim of this study was to investigate the effect of incubation on freezability of cholesterol loaded cyclodextrin (CLC) treated buffalo spermatozoa.
  
  
  METHODS
  Semen samples with mass motility of 3+ and greater, collected from Murrah buffalo bulls were utilized. Immediately after collection, four equal groups of semen sample were made. Group I was kept as control and diluted with Tris upto concentration of 60×10(6) sperm/ml, where as Groups II, III, and IV were treated with CLC at 3 mg/120× 10(6) spermatozoa, incubated at 37°C for action of CLC for 10, 15 and 20 min, respectively, and diluted with tris upto concentration of 60×10(6) sperm/ml. All groups were subjected to equilibration and freezing. The evaluation of semen samples from all groups was carried out at fresh, pre-freeze and post-thaw stage for progressive motility, viability and hypo-osmotic swelling response (HOS response).
  
  
  RESULTS
  At the pre-freeze stage, significantly (p<0.05) higher percentage of progressive motility and viability was observed in treatment groups as compared to control with no significant difference among treatment groups. HOS response was significantly (p<0.05) higher in treatment groups as compared to control at pre-freeze stage. At post-thaw stage, significantly (p<0.05) higher percentage of progressive motility, viability and HOS response was recorded in Group II as compared to control and other treatment groups (III and IV). Group II retained significant post-thaw motility and viability at various post-thaw incubation periods.
  
  
  CONCLUSIONS
  Incubation period of 10 min for CLC treated buffalo spermatozoa yielded significantly higher results in terms of freezability as compared to incubation for 15 and 20 min.
authors.affiliation: Division of Animal Reproduction, Gynecology & Obstetrics, National Dairy Research Institute, Karnal - 132001, Haryana, India.
authors.forename: S A
authors.initials: SA
authors.lastname: Lone
authors.affiliation: Germplasm Centre, Division of Animal Reproduction, Indian Veterinary Research Institute, Izatnagar, Bareilly - 243122, Uttar Pradesh, India.
authors.forename: J K
authors.initials: JK
authors.lastname: Prasad
authors.affiliation: Germplasm Centre, Division of Animal Reproduction, Indian Veterinary Research Institute, Izatnagar, Bareilly - 243122, Uttar Pradesh, India.
authors.forename: S K
authors.initials: SK
authors.lastname: Ghosh
authors.affiliation: Germplasm Centre, Division of Animal Reproduction, Indian Veterinary Research Institute, Izatnagar, Bareilly - 243122, Uttar Pradesh, India.
authors.forename: G K
authors.initials: GK
authors.lastname: Das
authors.affiliation: Germplasm Centre, Division of Animal Reproduction, Indian Veterinary Research Institute, Izatnagar, Bareilly - 243122, Uttar Pradesh, India.
authors.forename: B
authors.initials: B
authors.lastname: Balamurugan
authors.affiliation: Germplasm Centre, Division of Animal Reproduction, Indian Veterinary Research Institute, Izatnagar, Bareilly - 243122, Uttar Pradesh, India.
authors.forename: R
authors.initials: R
authors.lastname: Katiyar
authors.affiliation: Division of Livestock Economics and Statistics, Indian Veterinary Research Institute, Izatnagar, Bareilly - 243122, Uttar Pradesh, India.
authors.forename: M R
authors.initials: MR
authors.lastname: Verma
keywords: buffalo spermatozoa; cholesterol loaded cyclodextrin; incubation