Effect of Storage Time and Temperature of Equine Epididymis on the Viability, Motion Parameters, and Freezability of Epididymal Sperm

Monteiro, Gabriel Augusto; Guasti, Priscilla Nascimento; Rocha, Aline Silva; Martin, Ian; Sancler-Silva, Y. F. R.; Dell’Aqua, C.P.F.; Dell’Aqua, J.A.; Papa, Frederico Ozanam

doi:10.1016/j.jevs.2012.06.010

Cited by 31 publications

(20 citation statements)

References 15 publications

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“…Cooling between 4°C and 5°C reduces sperm metabolic activity, bacterial growth, and keeps sperm viable for long periods [13]. Similar results were observed during storage of the testis-epididymis complex at 5°C, which provided better preservation of epididymal sperm than storage at room temperature [21].…”

Section: Discussionsupporting

confidence: 66%

Viability of Pony Stallion Semen in Different Temperature and Dilution

Trentin

Rodrigues

Pessoa

et al. 2017

Acta Scientiae. Vet.

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Background: Artificial insemination and transport of cooled semen has been routinely used in equine industry in the past 20 years. However, more investigations are needed regarding the methods for long time storage in pony stallion semen. The effect of dilution and cooling temperature on pH, sperm motility, membrane integrity and mitochondrial activity were investigated before and after cooling of stallion semen.Materials, Methods & Results: Two ejaculates each from nine Brazilian ponies were diluted in a nonbuffered powder milk extender cooled at 5°C or 15°C for 48 h using three different dilutions (1:1, 1:2 or 1:3). Data were assessed by analysis of variance and the rate comparison was performed using the Duncan test. Samples diluted 1:1 at 5oC or 15°C showed higher pH values (7.63 ± 0.34 e 7.57 ± 0.27) and lower progressive motility (10.3 ± 11.05, 17.08 ± 9.95). All samples cooled at 15°C also showed lower incidence of morphologically altered spermatozoa (1:1 = 55.84%; 1:2 = 51.84%; 1:3 = 49.95%) [P < 0.01]. Mitochondrial activity was higher on the 1:3 dilution (0.86 ± 0.19 nm) at 5°C and on the 1:1 (0.89 ± 0.23 nm), 1:2 (0.93 ± 0.2 nm) and 1:3 (0.92 ± 0.2 nm) dilutions at 15°C. Progressive motility was higher when semen was diluted 1:3 and cooled at 15°C (42.22 ± 12.38; P < 0.05). Considering mitochondrial activity, similar results were observed when different dilutions of semen were used (P > 0.05) despite time and temperature. The pH, progressive motility, mitochondrial activity and membrane integrity remained similar (P > 0.05) on fresh semen samples independent of the dilution grade used. The best results were obtained when semen was diluted 1:3 and cooled at 15°C. All dilution grades were safe for fresh semen and pH wasincreased when semen was diluted and cooled for 48 h.Discussion: The methodology used to collect and process equine semen and semen from ponies is practically the same. Equine semen when sent for artificial insemination is usually cooled to 5°C. Our results showed that cooling reduces sperm viability, which has also been demonstrated by other studies. In contrast, the best cooling temperature was at 15°C. However, it is easier to keep the temperature at 5°C during transport, due to the large temperature oscillation that may occur during transportation. The semen of ponies can tolerate cooling at both 5 and 15°C. The 1:3 dilution cooled to 15°C provided better viability of pony sperm, and more stable pH during 48 h of cooling. Dilution 1:1 should not be used for cooling in powdered skim milk extender.

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Section: Discussionsupporting

confidence: 66%

Viability of Pony Stallion Semen in Different Temperature and Dilution

Trentin

Rodrigues

Pessoa

et al. 2017

Acta Scientiae. Vet.

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“…Another practical system for the transport of diluted semen that is also used in the field is Botutainer® (Botupharma, Botucatu, Brazil). According to the manufacturer, this system maintains the semen temperature at 5°C for up to 24 h. This transport box for cooled semen have been reported to be efficient in maintaining semen viable after cooling (Monteiro et al, 2013;Crespilho et al, 2014;Moscardini et al, 2014;Borges-Silva et al, 2016). Recently, programmable units for semen cryopreservation have been developed, which permit the selection of the desired cooling and freezing curve.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of cooling and freezing systems of bovine semen

Dias

Campanholi

Rossi

et al. 2018

Animal Reproduction Science

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Semen cryopreservation comprises different steps, among them are the cooling and freezing rates which significantly influence the quality of thawed sperm. Different systems with variable freezing rates are used for freezing bull semen in the field, with a consequence of variable success rates. The objective of this study was to compare different systems for freezing bull semen in the field. Five cooling methods of semen and two methods for the subsequent freezing phase (5 × 2 factorial scheme) were used. Two to four ejaculates were collected from 12 bulls with an electroejaculator. The ejaculates were diluted in BotuBov® to a concentration of 50 × 10 6 spermatozoa/mL in 0.5-mL straws. After dilution, the straws were cooled to 5°C in five cooling systems: TK 4000® at a cooling rate of −0.25°C/min (R1); TK 4000® at a rate of −0.5°C/min (R2); Minitube® refrigerator at a rate of −2.8°C/min (R3); Botutainer® at a rate of −0.65°C (R4), and domestic refrigerator at a rate of −2.0°C/min (R5). After stabilization at 5°C for 4 h, these straws were then submitted to two freezing systems: TK 4000® at a freezing rate of −15°C/min (C1) and Styrofoam box with liquid nitrogen at a rate of −19°C/min (C2). Sperm kinetics were evaluated by computer-assisted sperm analysis at four time points: in fresh semen, after cooling, post-thawing, and after the rapid thermal resistance test (TRT). In addition, plasma and acrosomal membrane integrity, mitochondrial potential and intracellular H 2 O 2 were analyzed after thawing by flow cytometry. The R1, R2 and R4 cooling systems were the most efficient in preserving sperm viability, membrane integrity and intracellular H 2 O 2. Samples frozen in the C1 system exhibited better post-thaw and post-TRT kinetics than C2 samples. In conclusion, slower cooling curves in conjunction with a constant freezing rate obtained with the programmable unit were more efficient for freezing bull semen in the field.

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“…Sperm maturation occurs in the tract of the epididymis, with the sperm acquiring important functions essential for fertilization such as motility, differentiation of the midpiece and acrosome [11] [12] and changes and modifications to the chromatin of the nucleus [5] [13] [31]. The differences detected in the sperm collected in different segments of the epididymis reflect the biochemical changes that occur during sperm passage [32] [33]. When sperm reach the cauda they come in contact with various substances including immobiline which greatly reduces their metabolism to minimize the energy demand and minimize the occurrence of residues; primarily because the epithelium has not been developed for absorbing wastes, which are higher in the segment of the head and somewhat less in the corpus of the epididymis [33] [34].…”

Section: Discussionmentioning

confidence: 99%

“…The differences detected in the sperm collected in different segments of the epididymis reflect the biochemical changes that occur during sperm passage [32] [33]. When sperm reach the cauda they come in contact with various substances including immobiline which greatly reduces their metabolism to minimize the energy demand and minimize the occurrence of residues; primarily because the epithelium has not been developed for absorbing wastes, which are higher in the segment of the head and somewhat less in the corpus of the epididymis [33] [34]. The greater concentration of sperm was obtained from the head of the epididymis, and was higher than that reported by others, e.g., James et al [35] and Guimarães et al [3].…”

Section: Discussionmentioning

confidence: 99%

Azteca Breed Horse Epididymal Sperm Evaluation: A Comparison of Head, Corpus and Cauda Sperm Quality

Parra-Forero¹,

Mendoza

Góngora³

et al. 2015

ARSci

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The main objective of this study was to determine differences in sperm recovered from the three segments of the epididymis, i.e., head, corpus and cauda, using routine measurements of sperm quality and DNA fragmentation assessment. There was a larger sperm concentration (P < 0.05) in the head (3.34 × 10 6 sperm/mL) compared with the corpus and cauda. The percentage of normal morphology was higher in the cauda but had lower motility (P < 0.05). The DNA fragmentation index was higher for sperm obtained from the head and body of the epididymis (P < 0.01) compared with the cauda. The DNA fragmentation index values were 12% higher (P < 0.01) using Wright stain compared with DAPI stain in sperm from the head and corpus of the epididymis, but similar in the cauda. Spermatobioscopy parameters match the maturation changes that occur in the epididymal tract with higher quality sperm being from the cauda, but with a lower concentration compared to sperm found in the head.

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Effect of Storage Time and Temperature of Equine Epididymis on the Viability, Motion Parameters, and Freezability of Epididymal Sperm

Cited by 31 publications

References 15 publications

Viability of Pony Stallion Semen in Different Temperature and Dilution

Viability of Pony Stallion Semen in Different Temperature and Dilution

Evaluation of cooling and freezing systems of bovine semen

Azteca Breed Horse Epididymal Sperm Evaluation: A Comparison of Head, Corpus and Cauda Sperm Quality

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