Laboratorio Of Andrologia

Zulia, Venezuela

Laboratorio Of Andrologia

Zulia, Venezuela
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Nava-Trujillo H.,Laboratorio Of Andrologia | Quintero-Moreno A.,Laboratorio Of Andrologia | Osorio-Melendez C.,Laboratorio Of Andrologia | Rubio-Guillen J.,Laboratorio Of Andrologia | And 2 more authors.
Revista Cientifica de la Facultad de Ciencias Veterinarias de la Universidad del Zulia | Year: 2011

The hypoosmotic swelling test (HOST) has been used to asses the sperm membrane functional integrity in bulls. However, for human, goat and dog semen, the water test (WT) has been proposed as a replacement to HOST, because WT uses distilled water and only five minutes of incubation. The aim of the present study was to compare the HOST and the WT to assess the sperm membrane functional integrity of cryopreserved bull semen. The percentage of sperm with swelling tail was higher in WT (54.85 ± 13.63%, P<0.0001) than HOST (42.90 ± 12.89%), but both were correlated positively (r = 0.77, PO.0001). In comparison with viability of thawed semen (52.52 ± 9.79%), only WT diminished significantly this parameter to 45.95 ± 8.69%, P<0.0001. Acrosome integrity of thawed semen (82.35 ± 8.21%) decreased significantly after HOST (57.87 ± 12.67%) and WT (39.00 ± 16.88%). Acrosome integrity in thawed semen was correlated positively with results after HOST (r = 0.52; P = 0.0126) and WT (r = 0.52; P = 0.0115). In conclusion, the WT can assess the sperm membrane functional integrity of cryopreserved bull semen and is able to replace the HOST.


Quintero-Moreno A.,Laboratorio Of Andrologia | Rubio-Guillen J.,Laboratorio Of Andrologia | Gonzalez-Villalobos D.,Laboratorio Of Andrologia | Madrid-Bury N.,Laboratorio Of Andrologia | And 2 more authors.
Revista Cientifica de la Facultad de Ciencias Veterinarias de la Universidad del Zulia | Year: 2011

The aim of this study was to investigate the effect of cryopreservation on the membrane integrity (MI) in the bull sperm, and determine the relationship between intact cryopreserved sperm and field fertility (FF). The eosin nigrosin exclusion and the hypo-osmotic swelling test were combined in a single test (HOS-EN test) to identify the spermatozoa with four types of MI. After cryopreservation, there was a marked decline (21.3%) in the percentage of spermatozoa with head membrane intact/ tail membrane intact (Type IV MI), and a significant increase in those with head membrane damaged/ tail membrane damaged (Type I MI) (P<0.001), whereas type II MI (head membrane intact /tail membrane damage) and type III MI (head membrane damage/ tail membrane intact) ocurred sparingly. A significant correlation was observed between the percentage of Type IV MI and FF (r=0.49, P<0.014). Cryopreservation causes a important demage in the plasmatic membrane of the spermatozoa. Intact MI is closely related to FF.


Quintero-Moreno A.,Laboratorio Of Andrologia | Nava J.,Laboratorio Of Andrologia | Osorio-Melendez C.,University of Zulia
Revista Cientifica de la Facultad de Ciencias Veterinarias de la Universidad del Zulia | Year: 2014

The objective of this study was to compare the sensitivity of three hyposmotic test (HOST) to evaluate the functional integrity of the plasmatic sperm membrane (IFMP) obtained from cryopreserved semen of 12 bulls. Were compared three solutions with different osmolarities (0,102 and 154 mOsm/kg) which were incubated at 37°C in a water bath. The solution with 0 mOsm/kg (A: ultra pure water) was incubated for five minutes (min), while of two remaining with different concentrations of sodium chloride were incubated for 30 min. Solution B and C comprise a hyposmotic media at 102 mOsm/kg and 154 mOsm/kg, respectively. Three samples were taken (100 μL) of thawed semen per bull, that were Introduced in vials containing 1000 μL of each hyposmotic solution. About smears performed with each medium hyposmotic and stained with eosin and nigrosin (EN), 100 spermatozoa were evaluated to determine (together) the percentages of viability and swelling flagellas (FR). The association of the HOST with EN was performed to identify the percentage of spermatozoas live or dead with FR or intacts. The results of the HOST demonstrated higher percentages of spermatozoes with FR for treatments B (49.75 ± 4.17) and C (56.33 ± 4.17) versus treatment A (32.16 ± 4.17) (P<0.05). These values indicate that the conditions B and C are optimal solutions, propitiating the helical twist of the flagellum, maintaining the IFMP of the sperm in 52% of cases. The osmolar condition A is not recommended because it causes loss of IFMP and cell death, thus was in evidence to appreciate that only 8% of sperm with flagella reacted survive after osmotic stress as low.


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.


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.

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