title: Effect of different semen extenders for the storage of chilled sperm in Tigrina (Leopardus tigrinus).
abstract: To enhance the conservation of endangered populations, the present study aimed to evaluate whether Tigrinas (Leopardus tigrinus) sperm could be conserved under refrigeration for short periods while maintaining sufficient quality for use in assisted-reproductive techniques (i.e., cryopreservation, in vitro fertilization). For this purpose, semen samples from 15 Tigrinas individuals were submitted to conventional and functional tests after different cooling periods (4 °C; 0, 12, and 24 hours postcooling), using TCM 199 (TCM), Ham's F10 (HAM), Ham's F10 with bovine serum albumin (HBSA), and Tris-citrate egg yolk (TEYC) extenders. In a second step, semen cooled using TEYC was supplemented with reduced glutathione (GSH) at different concentrations (0, 0.5, 1.0, and 1.5 mM). TEYC yielded superior results compared with TCM, HAM, and HBSA even after 24 hours of cooling in regard to the sperm motility index (SMI-TEYC: 50.2 ± 1.7%), high mitochondrial activity (TEYC: 51.4 ± 1.9%), plasma membrane integrity (TEYC: 53 ± 2.1%), and DNA integrity (TEYC: 56.3 ± 2.9%). In regard to the concentration of thiobarbituric-acid-reactive substances (TBARS), TEYC (1900.1 ± 341.4 ng/106spermatozoa) showed higher levels compared with the other extenders (HAM: 638.7 ± 121.6 ng/106spermatozoa; HBSA: 468.7 ± 95.6 ng/106spermatozoa; TCM: 169.6 ± 31.6 ng/106spermatozoa). However, GSH therapy had no effect. In conclusion, the TEYC extender may be useful in maintaining sperm parameters of Tigrinas for up to 24 hours at 4 °C. Furthermore, these results allow the transport of this material at a minimum quality to be further used for artificial insemination, in vitro fertilization, and the development of semen cryopreservation protocols.
authors.affiliation: Department of Animal Reproduction, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil.
authors.forename: D S R
authors.initials: DSR
authors.lastname: Angrimani
authors.affiliation: Department of Animal Reproduction, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil.
authors.forename: P M H
authors.initials: PMH
authors.lastname: Barros
authors.affiliation: Department of Animal Reproduction, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil.
authors.forename: J D A
authors.initials: JDA
authors.lastname: Losano
authors.affiliation: Department of Animal Reproduction, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil; Technology Institute of Paraná (TECPAR), Curitiba, Paraná, Brazil.
authors.forename: C N M
authors.initials: CNM
authors.lastname: Cortada
authors.affiliation: Division of Urology, Department of Surgery, Sao Paulo Federal University, Sao Paulo, Brazil.
authors.forename: R P
authors.initials: RP
authors.lastname: Bertolla
authors.affiliation: Department of Animal Reproduction, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil.
authors.forename: M A B V
authors.initials: MABV
authors.lastname: Guimarães
authors.affiliation: Department of Animal Reproduction, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil; Fundação Parque Zoológico de São Paulo (FPZSP), São Paulo, Brazil.
authors.forename: S H R
authors.initials: SHR
authors.lastname: Correa
authors.affiliation: Department of Animal Reproduction, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil.
authors.forename: V H
authors.initials: VH
authors.lastname: Barnabe
authors.affiliation: Department of Animal Reproduction, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil. Electronic address: mnichi@usp.br.
authors.forename: M
authors.initials: M
authors.lastname: Nichi
mesh_terms: D000818:Animals; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004530:Egg Yolk / Q000737:chemistry; D045989:Felidae / Q000502:physiology*; D005978:Glutathione / Q000494:pharmacology; D008297:Male; D012034:Refrigeration / Q000662:veterinary*; D012662:Semen Preservation / Q000379:methods / Q000662:veterinary*; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000187:drug effects / Q000502:physiology*
chemical_list: D003451:Cryoprotective Agents; D005978:Glutathione
keywords: Antioxidants; DNA fragmentation; Leopardus tigrinus; Reactive oxygen species; Short-term preservation