title: Quercetin reduces the in vitro production of H2O2 during chilled storage of rabbit spermatozoa.
abstract: Reactive oxygen species, such as hydrogen peroxide, H2O2, can reduce sperm quality during storage. This study evaluated the effect of methionine and quercetin on rabbit sperm quality during liquid storage over 96h. Semen was collected from adult bucks (n=4) and pooled following evaluation. In Experiment 1, pooled ejaculates were diluted with a Tris extender supplemented with methionine (1, 6 or 12mM), quercetin (50 or 200μM) or no antioxidant (control) and then subdivided for storage at 5°C or 15°C. Sperm quality was assessed by CASA (total motility [TM]) and flow cytometry (viability, acrosome integrity and H2O2 production) at 0, 48, 72 and 96h. Experiments were replicated three times. Motility was significantly higher in control samples and lowest following dilution with 200μM quercetin, irrespective of storage temperature. Storage at 15°C improved viability and acrosome integrity compared with 5°C, but produced significantly more H2O2 at 72 and 96h in sperm diluted with methionine or no antioxidant. Quercetin-supplemented spermatozoa exhibited lower levels of H2O2 at both storage temperatures for all incubation times (P<0.05). In Experiment 2, the concentration of quercetin (0, 25, 50, 100 and 200μM) was investigated with additional quality parameters; lipid peroxidation and DNA integrity. All concentrations of quercetin reduced H202 and lipid peroxidation during storage at 15°C, but were not beneficial for TM, viability, acrosome or DNA integrity. Only supplementation with 100 and 200μM quercetin resulted in similar H202 levels at 5°C and 15°C (P>0.05). Overall, quercetin-supplementation to sperm medium provided protection against oxidative stress in 15°C-stored rabbit spermatozoa over 96h.
authors.affiliation: Faculty of Veterinary Science, The University of Sydney, NSW 2006, Australia. Electronic address: danielle.johinke@sydney.edu.au.
authors.forename: D
authors.initials: D
authors.lastname: Johinke
authors.affiliation: Faculty of Veterinary Science, The University of Sydney, NSW 2006, Australia.
authors.forename: S P
authors.initials: SP
authors.lastname: de Graaf
authors.affiliation: Faculty of Veterinary Science, The University of Sydney, NSW 2006, Australia.
authors.forename: R
authors.initials: R
authors.lastname: Bathgate
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D002478:Cells, Cultured; D006861:Hydrogen Peroxide / Q000378:metabolism*; D008297:Male; D011794:Quercetin / Q000494:pharmacology*; D011817:Rabbits; D017382:Reactive Oxygen Species / Q000378:metabolism; D012034:Refrigeration / Q000662:veterinary; D055101:Semen Analysis / Q000662:veterinary; D012662:Semen Preservation / Q000009:adverse effects / Q000662:veterinary; D013094:Spermatozoa / Q000187:drug effects* / Q000378:metabolism*
chemical_list: D000975:Antioxidants; D017382:Reactive Oxygen Species; D011794:Quercetin; D006861:Hydrogen Peroxide
keywords: Antioxidant; Flow cytometry; Methionine; Oxidative stress; Quercetin; Rabbit