Modification of the short osmotic resistance test for evaluating the functional integrity of boar spermatozoa

Nascimento, Jenine; Nascimento, Evaldo; Melo, Maria Isabel Vaz de

doi:10.1590/s0102-09352001000300020

Cited by 2 publications

(8 citation statements)

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“…In classic ORT, the medium used is the hypotonic solution of 100mOsmol sodium citrate solution in one-hour incubation time. In SORT, the medium used were the hypotonic solution of 150mOsmol and75mOsmol sodium citrate solution with concentration of 1.5% and 0.75% respectively whereas in modified SORT, only 75mOsmol sodium citrate solution was used (Nascimento et al, 2001). The 10% form alinsaline solution was also used in both SORT and modified SORT tests (Nascimento et al, 2001).…”

Section: Medium For Sperm Preparationmentioning

confidence: 99%

“…In SORT, the medium used were the hypotonic solution of 150mOsmol and75mOsmol sodium citrate solution with concentration of 1.5% and 0.75% respectively whereas in modified SORT, only 75mOsmol sodium citrate solution was used (Nascimento et al, 2001). The 10% form alinsaline solution was also used in both SORT and modified SORT tests (Nascimento et al, 2001). It has been postulated that formaldehyde presence is able to exert a preservation of sperm in longer storage (El-Kon et al, 2009).…”

Section: Medium For Sperm Preparationmentioning

confidence: 99%

“…Slides were prepared at every interval in triplicates.The isosmotic solution (300mOsm) serves as control for classic ORT. For SORT test, the sperm suspension pellet was incubated with 300 mOsmol sodium-citrate solution followed by the addition of 50 µl 10% formalin saline performed inwater bath at 37 o C (Nascimento et al, 2001). Approximately 3µl of incubated test solution incubated was pippetted for slides preparation right after 5 minutes incubation has completed.…”

Section: Ort Sort and Modified Sort Solutionsmentioning

confidence: 99%

“…The extent of the viability of sperm incubated in ORT solutions is also important as after maximum tail coiling has occurred, sperm will start to die (Jeyendran et al 2009).Thus, this explains both tail coiling and acrosomal swelling as one of the parameters observed to determine functional membrane integrity of spermatozoa in ORT. Nascimento et al, (2001) introduced three approaches of ORT to test the functional membrane integrity of the sperm. These approaches were the classic ORT, short ORT (SORT) and modified SORT.The classic ORT is the traditional osmotic resistance test (ORT) that have a longer incubation time of one hour while SORT have an incubation time of 5 minutes and the modified SORT is a shorter version of SORT by eliminating one step in the SORT procedure.…”

Section: Introductionmentioning

confidence: 99%

“…These approaches were the classic ORT, short ORT (SORT) and modified SORT.The classic ORT is the traditional osmotic resistance test (ORT) that have a longer incubation time of one hour while SORT have an incubation time of 5 minutes and the modified SORT is a shorter version of SORT by eliminating one step in the SORT procedure. Nascimento et al (2001) uses boar sperm to test these three approaches where modified SORT was found to yield promising results. The shape of the sperm head in the boar is paddle shape as compared to mouse sperm which has a falciform head shape.…”

Section: Introductionmentioning

confidence: 99%

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Determination of the Osmotic Resistance Changes in Mouse Spermatozoa Using the Classic Osmotic Resistance Test, Short Osmotic Resistance Test and Modified Short Osmotic Resistance Test

Azmi¹,

Sukardi²

2018

theijst

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The acrosome is a very important structure on the sperm head. Made up of the outer and inner acrosomal membrane, the acrosome contained enzymes important to digest the zona pellucida of the ovum upon fertilization. Therefore, evaluation of the functional membrane integrity of the acrosome is one of the methods to recognize fertilizing capacity of the sperm. The method commonly used is the osmotic resistance test apart from the hypoosmotic test. Males can be better donors for sperm cryopreservation and this classification can be determine if osmotic resistance test showed excellent results. The traditional or classic osmotic resistance test (ORT) has a longer incubation time of two hours therefore a modified ORT was proposed with an incubation time of 5 minutes referred to as short osmotic resistance test (SORT), and later the modified SORT was introduced where samples were incubated in only 75 mOsmol solution for 5 minutes. As results vary for ejaculated sperm and cauda epididymal sperm, the aim of this study is to detect acrosomal membrane functional integrity in mouse sperm when incubated in classic ORT, short ORT and modified short ORT solutions. Cauda epididymal sperm from twenty mice were collected into Krebs Ringer's Buffer medium and divided into three parts: one for each test. Sperm were pooled, discarded of cell debris and centrifuged on a 2-step discontinuous Ficoll gradient. The sperm was collected from the pellet and incubated for 60 minutes in classic ORT buffer and 5 minutes in short ORT buffer and modified short ORT buffer. Tail swelling, proportion of normal acrosomes (%NAR) and viability was observed from smears taken at intervals of 10 minutes for classic ORT and 1 minute for SORT and modified SORT. Results were subjected to calculation for SORT and modified SORT only which is: SORT (5 min) =1/2[%NAR in 300mOsmol solution (5 min) + % NAR in 75mOsmol solution (5 min)] while modified SORT (5 min) =1/2[%NAR in samples from sperm morphology + % NAR in 75mOsmol solution (5 min)]. The results showed that classic ORT and modified SORT were found to yield the same findings however classic ORT and SORT showed different findings. The results suggest that the modified SORT can be apply to detect changes in acrosomal integrity to replace classic ORT as it is a rapid test and yields the same results as the classic ORT.

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Section: Medium For Sperm Preparationmentioning

confidence: 99%

Section: Medium For Sperm Preparationmentioning

confidence: 99%

Section: Ort Sort and Modified Sort Solutionsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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