Relationship among damaged chromatin, motility and viability in cryopreserved spermatozoa from Brahman bulls [Relación entre la alteración de la cromatina, movilidad y viabilidad en espermatozoides criopreservados de toros brahman]
Nava-Trujillo H.,Laboratorio Of Andrologia |
Quintero-Moreno A.,Laboratorio Of Andrologia |
Carruyo G.,University of Zulia |
Vilchez-Siu V.,University of Zulia |
And 3 more authors.
Revista Colombiana de Ciencias Pecuarias | Year: 2011
The aim of this study was to determine the percentage of sperm with damaged chromatic measure with toluidine blue stain and it ́s relationship with motility and viability in criopreserverd semen from Brahman bulls. Three ejaculates from six Brahman bulls were used. Immediately after thawing, sperms were stained with toluidine blue to establish chromatin integrity (sperms with normal chromatin were light blue or green while sperms with damaged chromatin were dark blue or violet). Sperms were also stained with eosin-nigrosin to determine viability (live sperms were unstained while dead sperms were pink). Motility was measured under light microscope. Effects of bull, ejaculate, and the interaction between variables were assessed. The percentage of live sperms was 50.02 (± 14.13%). The mean motility was 33.88 (± 12.43%), while the percentage of sperms with damaged chromatin was 4.17 (± 2.96%). Viability was positively correlated with motility (r=0.77217, p=0.0002), and negatively correlated with damaged chromatin sperms (r= -0.43104, p=0.0087). Motility percentage was negatively correlated with the percentage of sperms with damaged chromatin (r=-0.48337, p=0.0421). In conclusion, cryopreserved semen of Brahman bulls presented a low level of chromatin damage, and this trait was negatively correlated with sperm motility and viability.
Garcia-Peiro A.,Autonomous University of Barcelona |
Ribas-Maynou J.,Autonomous University of Barcelona |
Oliver-Bonet M.,Autonomous University of Barcelona |
Navarro J.,Autonomous University of Barcelona |
And 5 more authors.
BioMed Research International | Year: 2014
Varicocele is one of the most common causes of low semen quality, which is reflected in high percentages of sperm cells with fragmented DNA. While varicocelectomy is usually performed to ameliorate a patient's fertility, its impact on sperm DNA integrity in the case of subclinical varicocele is poorly documented. In this study, multiple DNA fragmentation analyses (TUNEL, SCD, and SCSA) were performed on semen samples from sixty infertile patients with varicocele (15 clinical varicoceles, 19 clinical varicoceles after surgical treatment, 16 subclinical varicoceles, and 10 subclinical varicoceles after surgical treatment). TUNEL, SCD, and SCSA assays all showed substantial sperm DNA fragmentation levels that were comparable between subclinical and clinical varicocele patients. Importantly, varicocelectomy did improve sperm quality in patients with clinical varicocele; however, this was not the case in patients with subclinical varicocele. In summary, although infertile patients with clinical and subclinical varicocele have similar sperm DNA quality, varicocelectomy should only be advised for patients with clinical varicocele. © 2014 Agustín García-Peiró et al.
Comparison between the sperm DNA fragmentation test using the sperm chromatin dispersion technique and the vitality index measured with the acridine orange test [Comparación entre el test de fragmentación de ADN espermático mediante la técnica de SCD y el índice de vitalidad medida con el test de naranja de acridina]
Ortega Lopez L.,Laboratorio Of Andrologia |
Vila E.O.,Laboratorio Of Andrologia |
Dominguez P.L.,Laboratorio Of Andrologia |
Segovia A.G.,Laboratorio Of Andrologia |
And 3 more authors.
Revista Internacional de Andrologia | Year: 2010
Introduction: Sperm DNA integrity is one of the most important factors currently analyzed in the study of male fertility. There are numerous publications on the utility of diverse methods analyzing sperm chromatin status. One of the simplest is the ethidium bromide/acridine orange (AO) test, which assesses the degree of sperm DNA denaturation according to its binding to acridine orange staining. However, the AO test is limited to these metachromatic properties and consequently provides limited information on the nuclear status of the sperm. Other techniques, such as the sperm chromatin dispersion (SCD) test, analyze sperm DNA fragmentation by assessing DNA strand breaks. Objective: To analyze whether there is a correlation between sperm vitality measured through the ethidium bromide/AO test and DNA fragmentation measured through the SCD test. In both cases, the samples were analyzed visually. Material and methods: Eighty-two semen samples from men undergoing fertility testing in the Clínica Tambre were included in this study. The ethidium bromide/AO test was used to test sperm vitality and the SCD test (Halosperm, Halotech) was used to evaluate sperm DNA fragmentation. The AO technique detects the proportion of dead sperm, depending on the degree nuclear DNA denaturation. Sperm with orange fluorescence indicate denatured DNA, while those with green fluorescence contain intact DNA.The percentage of fragmentation was measured by the improved SCD test (Halosperm). This test indicates DNA strand breaks based on the size of the halo obtained. In fragmented sperm, no halo is observed when the sample is examined with a fluorescence microscope and GelRed fluorochrome. Sperm with intact DNA show a halo.Student's t-test was used to compare the means between the two groups. Differences of p<0.05 were considered statistically significant. Results: The proportion of sperm with denatured DNA detected with the AO test was significantly correlated with the proportion of sperm DNA fragmentation detected with the SCD test (r = 0.5, P < 0.001). In the 82 samples analyzed, the mean vitality index with the AO test was 20.93% (6 ± 50) versus 28.35% (7.3 ± 40) for the fragmentation index. The proportion of sperm with denatured or fragmented DNA was also positively correlated with motility. The two assays showed a statistically significant negative correlation with sperm concentration. Discussion: This study demonstrates a correlation between the results of sperm vitality measured by the AO test and the degree of DNA fragmentation measured by the SCD test. These results suggest that the AO test could be the first option to determine sperm chromatin damage before more expensive or complicated tests are performed. © 2010 Sociedad Española de Andrología.