title: Methods for cryopreservation of guinea fowl sperm.
abstract: Conservation of indigenous poultry species is an important part of the new Hungarian agricultural strategy. Semen cryopreservation is the most practical method for the long term storage of poultry genetic material. The objective was to compare four protocols for cryopreservation of guinea fowl sperm (slow and fast programmable, freezing in nitrogen vapor, and pellet) and three cryoprotectants (10% ethylene glycol, 6% dimethyl-formamide and 6% dimethyl-acetamide). The efficiency of the methods was examined by in vitro tests (subjective motility scoring, sperm concentration, morphological and live/dead sperm analysis with eosin-aniline staining). Thereafter, the two most promising methods were tested by artificial insemination of frozen-thawed semen (3 times a week for 3 weeks using 300 million spermatozoa/hen), followed by candling of incubated eggs, assessment of fertilization, embryonic death, and hatching rate. The survival rate of live, intact spermatozoa was greatest (p≤0.05) in pellet method and the slow programmable protocol (with 10% ethylene glycol) (28.6 and 23.5%). The two best protocols (based on in vitro assessment of post-thaw semen quality) were subsequently tested in vivo with artificial insemination. The pellet method yielded a 64% fertility rate compared to slow protocol with only 30% fertility. Regardless, both freezing protocols significantly increased embryonic deaths compared to the control group (16,7; 9,1 and 8,3%, respectively). During the 3-week in vivo trial, fertility increased and early embryonic death decreased over time. According to the results the guinea fowl sperm could tolerate the fast freezing in pellet better than the slower freezing rates and resulted acceptable fertility rate.
authors: Institute for Small Animal Research and Co-ordination Centre for Gene Conservation, Gödöllő, Hungary. varadievi@hotmail.com
authors: Éva
authors: É
authors: Váradi
authors: Barbara
authors: B
authors: Végi
authors: Krisztina
authors: K
authors: Liptói
authors: Judit
authors: J
authors: Barna
mesh_terms: D000081:Acetamides / Q000737:chemistry; D000818:Animals; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000737:chemistry; D004126:Dimethylformamide / Q000737:chemistry; D004625:Embryo, Nonmammalian; D019855:Ethylene Glycol / Q000737:chemistry; D005260:Female; D005306:Fertilization; D046094:Galliformes / Q000502:physiology*; D007315:Insemination, Artificial / Q000662:veterinary*; D008297:Male; D055101:Semen Analysis / Q000662:veterinary*; D012662:Semen Preservation / Q000662:veterinary*; D013076:Sperm Count; D013081:Sperm Motility / Q000502:physiology; D013094:Spermatozoa / Q000502:physiology* / Q000648:ultrastructure
chemical_list: D000081:Acetamides; D003451:Cryoprotective Agents; D004126:Dimethylformamide; D019855:Ethylene Glycol; C013959:dimethylacetamide
references: J Reprod Fertil. 1966 Apr;11(2):257-62
references: 5949286
references: Br Poult Sci. 1995 Dec;36(5):805-11
references: 8746982
references: J Androl. 2002 Jan-Feb;23(1):107-13
references: 11780916
references: Theriogenology. 2003 Feb;59(3-4):875-88
references: 12517390
references: Acta Vet Hung. 2004;52(2):227-33
references: 15168754
references: Poult Sci. 1999 Apr;78(4):586-90
references: 10230914
references: Anim Reprod Sci. 2000 Jul 2;60-61:481-92
references: 10844218
references: Anim Reprod Sci. 2011 Nov;129(1-2):89-95
references: 22078301
references: Poult Sci. 1986 Jan;65(1):1-15
references: 3960808
references: Poult Sci. 2011 Sep;90(9):2047-53
references: 21844272
references: J Androl. 2003 Jul-Aug;24(4):621-8
references: 12826702
references: Poult Sci. 2006 Feb;85(2):216-22
references: 16523616
references: Theriogenology. 2011 Sep 15;76(5):794-801
references: 21664665
references: Poult Sci. 2007 Mar;86(3):555-64
references: 17297169
references: Reproduction. 2005 Mar;129(3):371-8
references: 15749963
references: Hum Reprod. 2002 Apr;17(4):990-8
references: 11925396
references: Theriogenology. 2002 Jul 1;58(1):19-27
references: 12182361
references: Poult Sci. 2006 Feb;85(2):210-5
references: 16523615