Establishing the hypoosmotic swelling test for sperm analysis in collared peccaries (Pecari tajacu)

Santos, E.A.A.; Sousa, Patrícia Cunha; Peixoto, Gislayne Christianne Xavier; Simão, Bruno Rodrigo; Oliveira, Moacir Franco de; Silva, Alexandre Rodrigues

doi:10.1590/s0102-09352013000400044

Cited by 23 publications

(14 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Assessment of the plasma membrane functional integrity was performed by the hyposmotic swelling test (HOST) by adding 10µL of semen to 90µL distilled water (0 mOsmol/L) followed by immersion in dry bath at 37°C for 40 min (Santos et al 2013). After incubation, reading of percentage of HOST reactive sperm was performed under optical microscopy (Olympus  CX 21), using immersion objective (obj.1000x) in wet mount between slide and coverslip stained with 3% rose bengal.…”

Section: Methodsmentioning

confidence: 99%

Use of the ACP® and BTS extenders for cooling at 15°C white-lipped peccary (Tayassu pecari) semen

et al. 2019

View full text Add to dashboard Cite

Knowledge about reproduction of white-lipped peccary is of great importance to assist with the conservation of this species and enable its rational use in captivity. This study aimed to evaluate the effect of ACP-103®, ACP-116® and BTS semen extenders on sperm viability during cooling of Tayassu pecari semen. Five ejaculates from four adult males were chilled. The animals were submitted to the protocols of sedation and anesthesia for semen collection by the electroejaculation method. After collection, the semen was macro- and microscopically assessed and diluted to reach 35x106 spermatozoa/mL in each of the three different extenders tested. The fresh-extended semen was packed in a BotuFLEX® thermal box to keep samples at 15°C for 24 hours. After cooling, the following semen parameters were analyzed: sperm motility, functional and structural integrity of sperm membranes, mitochondrial activity, chromatin condensation, and the thermoresistance test was performed. The parameters sperm motility, structural and functional integrity of sperm membranes, mitochondrial activity, and chromatin condensation were preserved after use of the extenders tested, and were similar to those of in natura semen (p>0.05). Curvilinear velocity (VCL) (p<0.05) was the only parameter with reduced values after cooling regardless of the extender used. The percentage of sperm with normal morphology was greater in samples cooled using the BTS extender (p<0.05). The ACP-103®, ACP-116® and BTS extenders can be used for the cooling and preservation of white-lipped peccary semen at 15°C for 24 hours.

show abstract

Section: Methodsmentioning

confidence: 99%

Use of the ACP® and BTS extenders for cooling at 15°C white-lipped peccary (Tayassu pecari) semen

et al. 2019

View full text Add to dashboard Cite

show abstract

“…The functional integrity of the sperm membrane was verified by evaluating the sperm osmotic response under a hypo‐osmotic swelling test, using distilled water (0 mOsm/L) as the hypo‐osmotic solution. A total of 200 cells were examined, and those with a swollen coiled tail were considered as presenting a functional membrane (Santos et al, ).…”

Section: Methodsmentioning

confidence: 99%

Ultrastructural description of fresh and frozen/thawed sperm derived from collared peccaries (Pecari tajacu Linnaeus, 1,758)

Bezerra

Souza

Silva

et al. 2018

Microscopy Res & Technique

View full text Add to dashboard Cite

The aim was to describe, through scanning electron microscopy (SEM) and transmission electron microscopy (TEM), the ultrastructure of peccaries' fresh and frozen-thawed sperm. For that, semen derived from three mature males was obtained by electroejaculation and evaluated for motility, membrane integrity, membrane functionality, chromatin integrity, and morphology through light microscopy. Samples were frozen using a Tris extender plus egg yolk (20%) and glycerol (6%). Then, fresh and frozen-thawed semen samples were mixed in different sperm pools that were processed for SEM and TEM. Sperm motility, membrane integrity, and functionality were impaired (p < .05) by freezing-thawing procedures, but sperm morphology, and chromatin integrity evaluated by light microscopy were not significantly affected. The SEM revealed that peccaries' sperm presents a flattened head in a paddle format, measuring 6.07 μm in length and 3.84 μm in width, with a vastus acrosome (4.46 μm). Normal tails measure 38.11 μm, being formed by an extensive midpiece with 15.52 μm in length. In frozen-thawed samples, both SEM and TEM provide us information about damage undetected through light microscopy as the presence of vesicles in the acrosome, loose plasma membrane, vacuolized mitochondria, dense fibers disorganized, and decondensed chromatin. In conclusion, we provide the first description of the sperm ultrasctruture in collared peccaries. Moreover, SEM and TEM help us to identify some nanometric damage provoked by freezing-thawing procedures, thus providing valuable information for the improvement of such important protocols used for biobanking formation. K E Y W O R D Sfreezing-thawing, morphology, scanning electron microscopy, semen, Tayassu tajacu, transmission electron microscopy

show abstract

“…Functional membrane integrity was evaluated as the sperm osmotic response to a hypoosmotic swelling (HOS) test, using distilled water (0mOsm/l). Two hundred sperm were examined and those with a swollen, coiled tail were considered to have a functional membrane (Santos et al ., 2013). Sperm viability was determined by bromophenol blue stained smears, using light microscopy (×400 magnification), based on analyses of 200 cells per slide (Sousa et al , 2013).…”

Section: Methodsmentioning

confidence: 99%

Cryopreservation of collared peccary (Pecari tajacu L., 1758) epididymal sperm using extenders based on Tris and powdered coconut water (ACP®-116c)

Bezerra

Silva

Sousa

et al. 2018

Zygote

View full text Add to dashboard Cite

SummaryThe aim of this study was to establish a functional freezing-thawing protocol for epididymal sperm of collared peccaries (Pecari tajacu L., 1758) by comparing different extenders. The epididymal sperm from 12 sexually mature males was recovered by retrograde flushing using Tris-based or coconut water-based (ACP®-116c) extenders. After initial evaluation, samples were diluted and frozen with the same extenders to which 20% egg yolk and 6% glycerol were added. After 2 weeks, thawing was performed at 37°C/60 s and sperm motility, vigour, morphology, functional membrane integrity, sperm viability, sperm plasma membrane integrity, and a computer-assisted semen analysis (CASA) were assessed. In addition, to evaluate the survival of frozen-thawed sperm, a thermal resistance test (TRT) was executed. Samples preserved using Tris were in better condition compared with those preserved using ACP®, showing higher values for most assessments performed, including CASA and the TRT (P<0.05). After determining Tris to be the better of the two extenders, additional samples were thawed using different thawing rates (37°C/60 s, 55°C/7 s, 70°C/8 s). Sperm thawed at 37°C/60 s had the greatest preservation (P<0.05) of viability (54.1 ± 5.9%) and functional membrane integrity (43.2 ± 5.4%), and had higher values for various CASA parameters. In conclusion, we suggest the use of a Tris-based extender added to egg yolk and glycerol for the cryopreservation of epididymal sperm obtained from collared peccaries. In order to achieve better post-thawing sperm quality, we suggest that samples should be thawed at 37°C/60 s.

show abstract

Establishing the hypoosmotic swelling test for sperm analysis in collared peccaries (Pecari tajacu)

Abstract: Communication [Comunicação] Establishing the hypoosmotic swelling test for sperm analysis in collared peccaries (Pecari tajacu)[Estabelecimento do teste hiposmótico para a análise de sêmen de catetos (Pecari tajacu)]

Cited by 23 publications

References 10 publications

Use of the ACP® and BTS extenders for cooling at 15°C white-lipped peccary (Tayassu pecari) semen

Use of the ACP® and BTS extenders for cooling at 15°C white-lipped peccary (Tayassu pecari) semen

Ultrastructural description of fresh and frozen/thawed sperm derived from collared peccaries (Pecari tajacu Linnaeus, 1,758)

Cryopreservation of collared peccary (Pecari tajacu L., 1758) epididymal sperm using extenders based on Tris and powdered coconut water (ACP®-116c)

Contact Info

Product

Resources

About