Effects of matrix filtration of low-quality boar semen doses on sperm quality.
The aim of this work was to develop a method to enhance the sperm parameters of ejaculates with low sperm quality from Piétrain boars. Seminal doses were filtered through columns of DEAE Sephadex (length 2.5 +/- 0.5 cm), CM Sephadex (length 5 +/- 0.5 cm), glass wool (length 2 +/- 0.5 cm) or glass bead (length 10 +/- 0.5 cm), with an exit flow rate of 1 ml/40 s in all cases. For each male, 10 ml of the sperm cell-rich fraction diluted at 1 : 6 were filtered. Sperm quality was assessed before and after filtration. Sperm morphology, sperm motility and sperm concentration were determined using the computer program sca((R)) 2002 Production, and sperm viability was evaluated by fluorescence multistaining. Osmotic resistance test and hyperosmotic resistance test were used to determine the osmotic resistance of spermatozoa, whereas l-lactate production estimated the metabolic activity. Results showed a decrease of sperm concentration and osmotic resistance of spermatozoa after filtration in the four matrixes. However, an increase in the frequency of viable spermatozoa with intact acrosome after filtration in glass bead columns and an increase of morphologically normal spermatozoa after filtration in Sephadex CM-50, glass wool and glass bead columns were observed. Despite the decrease in the frequency of progressive motile spermatozoa, l-lactate production and mitochondrial sheath integrity maintained constant after filtration. Our findings indicate that column filtration is an effective method to enhance the sperm quality by selecting viable and morphologically normal spermatozoa without altering DNA, plasma membrane, mitochondrial sheath integrity or inducing premature acrosome reaction.
Bussalleu
E
E
Biotecnologia de la Reproducció Animal i Humana, Departament de Biologia, Facultat de Ciències, Universitat de Girona, Girona, Catalonia, Spain. eva.bussalleu@udg.edu
Pinart
E
E
Rivera
M M
MM
Briz
M
M
Sancho
S
S
Yeste
M
M
Casas
I
I
Fàbrega
A
A
Rigau
T
T
Rodriguez-Gil
J E
JE
Bonet
S
S
D000818:Animals; D002469:Cell Separation / Q000379:methods / Q000662:veterinary*; D002470:Cell Survival; D002845:Chromatography / Q000662:veterinary; D002852:Chromatography, Ion Exchange / Q000662:veterinary; D005374:Filtration / Q000379:methods / Q000662:veterinary*; D005898:Glass; D008297:Male; D008863:Microspheres; D012661:Semen / Q000166:cytology; D013076:Sperm Count; D013081:Sperm Motility; D013094:Spermatozoa / Q000002:abnormalities / Q000166:cytology / Q000502:physiology*; D013552:Swine