title: Use of a novel washing method combining multiple density gradients and trypsin for removing human immunodeficiency virus-1 and hepatitis C virus from semen.
abstract: OBJECTIVE
  To determine the effectiveness of a novel treatment designed to remove human immunodeficiency virus (HIV) -1 and hepatitis C virus (HCV) from spiked semen and to evaluate sperm motility and viability after treatment.
  
  
  METHODS
  A prospective clinical laboratory-based study.
  
  
  METHODS
  The human studies were conducted in academic and national research environments. The bovine study was conducted in an accredited research facility.
  
  
  METHODS
  Healthy volunteers provided the semen samples used in the human studies; abattoir-derived material was used for the bovine embryo production study.
  
  
  METHODS
  None.
  
  
  METHODS
  Cytopathic, reverse transcriptase-polymerase chain reaction, and branched DNA assays were used to test the efficacy of the procedure for inactivating or removing viruses from spiked semen; standard semen evaluation criteria were used to assess the effects of the procedures on sperm motility and viability.
  
  
  RESULTS
  Trypsin exposure significantly reduced the infectivity of HIV-1. The triple density gradient treatment, with or without trypsin, had no detrimental affect on fresh or cryopreserved/thawed sperm 2-48 hours after treatment. The treatment of semen spiked with HIV-1 or HCV indicated that the procedure was effective for reducing viral copies to undetectable levels or below levels of clinical relevance.
  
  
  CONCLUSIONS
  The procedure was effective for significantly inactivating or reducing HIV-1 and HCV in spiked semen without adversely affecting sperm quality.
authors.affiliation: The Bill and Berniece Grewcock Center for Conservation and Research, Omaha, Nebraska 68107, USA. NaidaL@omahazoo.com
authors.forename: Naida M
authors.initials: NM
authors.lastname: Loskutoff
authors.forename: Carin
authors.initials: C
authors.lastname: Huyser
authors.forename: Raksha
authors.initials: R
authors.lastname: Singh
authors.forename: David L
authors.initials: DL
authors.lastname: Walker
authors.forename: Alan R
authors.initials: AR
authors.lastname: Thornhill
authors.forename: Lynn
authors.initials: L
authors.lastname: Morris
authors.forename: Lynne
authors.initials: L
authors.lastname: Webber
mesh_terms: D000818:Animals; D002417:Cattle; D002499:Centrifugation, Density Gradient / Q000379:methods; D003718:Denmark; D005260:Female; D015497:HIV-1 / Q000187:drug effects* / Q000502:physiology; D016174:Hepacivirus / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D008297:Male; D011446:Prospective Studies; D011792:Quebec; D012661:Semen / Q000187:drug effects* / Q000502:physiology / Q000821:virology*; D013081:Sperm Motility / Q000187:drug effects* / Q000502:physiology; D014357:Trypsin / Q000494:pharmacology*; D019562:Viral Load / Q000706:statistics & numerical data
chemical_list: D014357:Trypsin