""The development of new procedures aimed at improving the quality of ram cryopreserved semen is a suitable goal to be pursued. In previous research was showed that the He-Ne laser irradiation can biostimulates sperm motility in different animal species (Ocaña-Quero et al. 1997; Ruiz-Pesini et al. 2000; Corral-Baquès et al., 2008; Iaffaldano et al., 2005; 2010). However, the effect of He-Ne laser was never investigated, to the best of our knowledge, in severely stressed semen, as in the freezing\\\/thawing procedure and, in particular, in ram semen.. Therefore, the aim of this study was to investigate whether and how two energy doses of laser irradiation (3.96 and 6.12 J\\\/cm2) can improve the qualitative characteristics and energetic profiles of ram spermatozoa after freezing-thawing process.. Cryopreserved ram sperm from “merinizzata italiana” breed was kindly purchased by “APA” association. Samples were thawed in a water bath at 37°C for 30 seconds. Each pool was divided into three aliquots: one was kept as a control and the others two were irradiated with He–Ne laser (wavelenght 632.8 nm; 6mW at two different energy dose (3.96 and 6.12 J\\\/cm2). The total mass. and forward progressive motility (subjectively assessed by light microscopy), viability (SyBr-PI staining), DNA intactness (Acridine orange staining), osmotic-resistance (HOS-test) and acrosome integrity (FITC-PSA) were evaluated after irradiation.. In parallel, the effect of irradiation on biochemical parameters of samples was evaluated by measuring the activity of cytochrome oxidase (COX) and the energetic charge (HPLC).. In all cases we found that the lower dose of laser energy resulted in a decrease in both mass and progressive motility, but not in viability, acrosome and DNA integrity; in parallel, this energy dose resulted in a stimulation of cytochrome oxidase (COX). On the contrary, the higher energy dose slightly increased progressive motility, viability, osmotic resistance and acrosome integrity without affecting mass motility and DNA integrity; energy charge and COX activity were found not to differ with respect to the control. Further studies are needed to confirm these results with a larger number of samples and to evaluate the effect of laser irradiation utilizing higher energy doses.. ""
He-Ne laser irradiation effects on cryopreserved ram semen.
IAFFALDANO, Nicolaia;DI IORIO, Michele;ROCCO, MARTINA;MANCHISI, Angelo;PAVENTI, Gianluca;
2013-01-01
Abstract
""The development of new procedures aimed at improving the quality of ram cryopreserved semen is a suitable goal to be pursued. In previous research was showed that the He-Ne laser irradiation can biostimulates sperm motility in different animal species (Ocaña-Quero et al. 1997; Ruiz-Pesini et al. 2000; Corral-Baquès et al., 2008; Iaffaldano et al., 2005; 2010). However, the effect of He-Ne laser was never investigated, to the best of our knowledge, in severely stressed semen, as in the freezing\\\/thawing procedure and, in particular, in ram semen.. Therefore, the aim of this study was to investigate whether and how two energy doses of laser irradiation (3.96 and 6.12 J\\\/cm2) can improve the qualitative characteristics and energetic profiles of ram spermatozoa after freezing-thawing process.. Cryopreserved ram sperm from “merinizzata italiana” breed was kindly purchased by “APA” association. Samples were thawed in a water bath at 37°C for 30 seconds. Each pool was divided into three aliquots: one was kept as a control and the others two were irradiated with He–Ne laser (wavelenght 632.8 nm; 6mW at two different energy dose (3.96 and 6.12 J\\\/cm2). The total mass. and forward progressive motility (subjectively assessed by light microscopy), viability (SyBr-PI staining), DNA intactness (Acridine orange staining), osmotic-resistance (HOS-test) and acrosome integrity (FITC-PSA) were evaluated after irradiation.. In parallel, the effect of irradiation on biochemical parameters of samples was evaluated by measuring the activity of cytochrome oxidase (COX) and the energetic charge (HPLC).. In all cases we found that the lower dose of laser energy resulted in a decrease in both mass and progressive motility, but not in viability, acrosome and DNA integrity; in parallel, this energy dose resulted in a stimulation of cytochrome oxidase (COX). On the contrary, the higher energy dose slightly increased progressive motility, viability, osmotic resistance and acrosome integrity without affecting mass motility and DNA integrity; energy charge and COX activity were found not to differ with respect to the control. Further studies are needed to confirm these results with a larger number of samples and to evaluate the effect of laser irradiation utilizing higher energy doses.. ""I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
