The Influence of Cigarette Smoking on Sperm Quality and Sperm Membrane Integrity

Routine methods for the evaluation of sperm quality are not sufficiently useful to determine the sensitivity of sperm cells to cold shock. The aim of our preliminary study was to determine whether the sperm plasma membrane integrity evaluated by modified hypoosmotic swelling test based on simple hypoosmotic swelling test (HOS test) and eosin-nigrosin staining could be helpful in predicting the degree of boar sperm survivability during semen cryopreservation. Ejaculates collected from 24 boars and 20 bulls were used in the experiment. Fresh ejaculates were evaluated by routine sperm analysis and a modified HOS test, and subsequently frozen. Sperm cryosurvivability was defined as the percentage of motile spermatozoa that survived the freezing process. A higher percentage of sperm was recovered after the thawing of semen with a higher percentage of HOS-positive and eosin-negative sperm (P < 0.01). Both indicators were found to be correlated (r = 0.707 and r = 0.705, respectively; P < 0.01). Moreover, the percentage of HOS-positive and eosin-negative sperm was similar to the percentage of viable sperm after thawing as determined by traditional eosin-nigrosin staining in boars (50.90 ± 9.88% and 49.31 ± 11.63%, respectively) and bulls (55.91 ± 10.34% and 55.63 ± 6.64%, respectively) and both indicators showed a positive correlation (r = 0.558 and r = 0.504, respectively; P < 0.05). In conclusion, based on the obtained results, we can assume that the modified HOS test can detect differences in sperm membrane resistance which allows assessment of semen quality from the perspective of its further use, e.g. cryopreservation.

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Efektivitas Penambahan Vitamin E (alfa-Tokoferol) dalam Medium Pencucian Sperma dengan Sentrifugasi terhadap Kualitas Spermatozoa Sapi Brahman

Jurnal Agripet ◽

10.17969/agripet.v12i2.196 ◽

2012 ◽

Vol 12 (2) ◽

pp. 7-13

Author(s):

Dasrul Dasrul ◽

Rasmaidar Rasmaidar ◽

Abdul Harris

Keyword(s):

Vitamin E ◽

Sperm Quality ◽

Membrane Integrity ◽

Average Percentage ◽

Treatment Groups ◽

Frozen Semen ◽

Brahman Cattle ◽

Sperm Membrane ◽

Better Than

Effect of vitamin E addition (alfa-tokoferol) into sperm washing medium by centrifuge on the quality of Brahman cattle spermatozoa ABSTRACT. The aims of study to determine the effectiveness of the addition of vitamin E in the washing medium by centrifugation on sperm quality Brahman cattle. frozen semen of Brahman cattle, divided into 4 treatment groups addition of vitamin E in the washing medium: 0.0gr/100 ml medium (K0), 0.1gr /100 ml medium (K1); 0.2gr/100 ml medium (K2) and 0.3 g / 100 ml medium (K4), each group was repeated 5 times. Examination of motility, viability and integrity of sperm membrane performed according to WHO standards. The data obtained were analyzed with one-way ANOVA and Duncan test. The average percentage of motility, viability and membrane integrity of spermatozoa in the addition of vitamin E were significantly different (P 0.05) compared to the control. Percentage of motility, viability and membrane integrity of spermatozoa in the group K2 significantly higher (P 0.05) compared with the group K3: K1 and K0. Percentage of motility, viability and sperm capacitation and sperm live on group K3 significantly higher (P 0.05) compared with the K1 and K0. While the percentage motility of spermatozoa in the group K1 higher were not significant (P 0.05) compared with the group K0. The addition of vitamin E in the medium on the process of washing spermatozoa Brahman cattle. The addition of vitamin E 0.2gr/100ml better than vitamin E 0.1gr/100ml and 0.3gr/100ml in maintaining the percentage of motility and live spermatozoa Brahman cattle.

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14 Sperm membrane integrity in ejaculates from young bulls may be improved by single-layer centrifugation

Reproduction Fertility and Development ◽

10.1071/rdv31n1ab14 ◽

2019 ◽

Vol 31 (1) ◽

pp. 133

Author(s):

E. Hurri ◽

A. Johannisson ◽

I. Lim-Verde ◽

J. M. Morrell

Keyword(s):

Sperm Quality ◽

Pearson Correlation ◽

Animal Health ◽

Sperm Concentration ◽

Membrane Integrity ◽

Single Layer ◽

Before And After ◽

Sperm Membrane ◽

Chromatin Integrity ◽

Species Production

Sperm quality in the first ejaculates of young bulls is often poor, leading to rejection of these samples for commercial freezing. It may be several weeks before sperm quality reaches the thresholds regarded as being acceptable for commercial production. Single-layer centrifugation (SLC) has been shown to separate robust bull spermatozoa from the rest of the ejaculate (Goodla et al. 2015 J. Dairy Sci. 97, 2204-2212; Nongbua et al. 2017 Reprod Domest. Anim. 52, 596-602). The aim of this study was to test whether SLC can be used to select spermatozoa from the ejaculates of young bulls and thus freeze samples earlier than is currently possible. Ejaculates (at least 3 per bull; n=33) were collected from 9 young Holstein or Swedish Red bulls, 255 to 415 days old. The sperm concentration was adjusted to 69×106 spermatozoa mL−1 with Andromed™ (Minitub, Tiefenbach, Germany) before layering part of the sample over 4mL of Bovicoll colloid. The remaining sample served as the unselected control. After centrifugation at 300×g for 20min, the sperm pellet was aspirated into fresh extender. Both control (uncentrifuged) and SLC samples were frozen and stored in LN pending analysis. Membrane integrity was evaluated by flow cytometry after staining for 10min at 37°C with SYBR14/propidium iodide (Live-Dead Sperm Viability Kit, L-7011; Invitrogen, Carlsbad, CA, USA). Paired t-test was used to compare results for control and SLC samples, and Pearson correlation was used for age and membrane integrity. The median age at which the samples were taken was 319 days (range: 255-415 days). Membrane integrity varied considerably among bulls (range: 11-72%). The mean proportion of membrane-intact spermatozoa in controls and SLC samples was 40±15 and 47±16%, respectively (P<0.01). In 21 of the 33 ejaculates (64%), the SLC sample had a higher membrane integrity than the controls (Table 1). Age in days was positively correlated with membrane integrity for SLC samples (r=0.40; P<0.05) but not for controls. These results suggest that SLC might be a useful technique for selecting membrane-intact spermatozoa from the ejaculates of young bulls. However, there is considerable variation among bulls, possibly due to age, which is currently being investigated. Other parameters of sperm quality, such as chromatin integrity, mitochondrial membrane potential reactive oxygen species production, and motility, are also being evaluated. Table 1.Age and membrane integrity (mean±standard deviation) in ejaculates from young bulls before and after SLC (n=33) This project was funded by the Royal Swedish Academy of Agriculture and Forestry, Viking Genetics, and the Faculty for Veterinary Medicine and Animal Health, SLU. We thank Viking Genetics for supplying the semen samples.

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Freezability of Andalusian donkey (Equus asinus) spermatozoa: effect of extenders and permeating cryoprotectants

Reproduction Fertility and Development ◽

10.1071/rd14449 ◽

2016 ◽

Vol 28 (12) ◽

pp. 1990 ◽

Cited By ~ 15

Author(s):

D. Acha ◽

M. Hidalgo ◽

I. Ortiz ◽

M. J. Gálvez ◽

J. J. Carrasco ◽

...

Keyword(s):

Sperm Motility ◽

Sperm Quality ◽

Membrane Integrity ◽

Egg Yolk ◽

Equus Asinus ◽

Sperm Membrane ◽

Freeze Test ◽

Pooled Samples

The aim of this study was to compare the effect of two semen extenders and four permeating cryoprotectants on post-thaw sperm quality of Andalusian donkeys. First, 32 ejaculates were pooled, split and frozen in either Gent B or INRA 96 with egg yolk and glycerol. Second, 12 pooled semen samples were simultaneously frozen in Gent B (glycerol) or Gent A containing ethylene glycol (EG; 1 or 1.5%) or dimethyl sulfoxide (DMSO; 1.5 or 2%). Finally, nine pooled samples were simultaneously cryopreserved in Gent A containing 1% EG (as control), dimethylformamide (DMFA; 1 or 2.5%) or a combination of 1% EG and 1.5% DMFA. Gent B yielded a higher (P < 0.01) post-thaw sperm motility than modified INRA96. EG 1% increased the sperm membrane integrity (P < 0.001), whereas DMSO affected sperm motility and membrane integrity (P < 0.001). DMFA 2.5% yielded higher (P < 0.001) values for sperm motility and membrane integrity. We concluded that Gent B improves in vitro post-thaw sperm quality of donkey spermatozoa, but the replacement of glycerol with 1% EG or 2.5% DMFA increased sperm protection against cryodamage. The use of DMSO for freezing donkey semen was unsuccessful and a toxic effect is suspected. These extenders should be included in the pre-freeze test for each donkey.

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Aquaporins 7 and 11 in boar spermatozoa: detection, localisation and relationship with sperm quality

Reproduction Fertility and Development ◽

10.1071/rd14237 ◽

2016 ◽

Vol 28 (6) ◽

pp. 663 ◽

Cited By ~ 17

Author(s):

Noelia Prieto-Martínez ◽

Ingrid Vilagran ◽

Roser Morató ◽

Joan E. Rodríguez-Gil ◽

Marc Yeste ◽

...

Keyword(s):

Western Blot ◽

Sperm Motility ◽

Western Blotting ◽

Water Permeability ◽

Mammalian Cells ◽

Sperm Quality ◽

Critical Role ◽

Membrane Integrity ◽

Sperm Membrane ◽

Boar Spermatozoa

Aquaporins (AQPs) are integral membrane water channels that allow transport of water and small solutes across cell membranes. Although water permeability is known to play a critical role in mammalian cells, including spermatozoa, little is known about their localisation in boar spermatozoa. Two aquaporins, AQP7 and AQP11, in boar spermatozoa were identified by western blotting and localised through immunocytochemistry analyses. Western blot results showed that boar spermatozoa expressed AQP7 (25 kDa) and AQP11 (50 kDa). Immunocytochemistry analyses demonstrated that AQP7 was localised in the connecting piece of boar spermatozoa, while AQP11 was found in the head and mid-piece and diffuse labelling was also seen along the tail. Despite differences in AQP7 and AQP11 content between boar ejaculates, these differences were not found to be correlated with sperm quality in the case of AQP7. Conversely, AQP11 content showed a significant correlation (P < 0.05) with sperm membrane integrity and fluidity and sperm motility. In conclusion, boar spermatozoa express AQP7 and AQP11, and the amounts of AQP11 but not those of AQP7 are correlated with sperm motility and membrane integrity.

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Relationship of extender and packaging system an the length of preservation and the quality of chilled semen of Boer goat

Jurnal Ilmu Ternak dan Veteriner ◽

10.14334/jitv.v21i1.1350 ◽

2016 ◽

Vol 21 (1) ◽

pp. 49

Author(s):

Arie Febretrisiana ◽

. Anwar ◽

Simon Sinulingga

Keyword(s):

Sperm Motility ◽

Sperm Quality ◽

Membrane Integrity ◽

Egg Yolk ◽

Packaging System ◽

Boer Goat ◽

Sperm Membrane ◽

Relationship Of ◽

Artificial Vagina

<p class="abstrak2">The aim of this research was to compare the effectiveness of different extender (either Triladyl or Tris Egg Yolk extender) and different packaging method (pool and straw) of chilled semen an the length of preservation and the quality of chilled semen of Boer goat. Semen was collected using an artificial vagina from 3 two years old Boer bucks with body weight of 50-55 kg. It was evaluated under a microscope, then each was diluted either in Tris egg yolk extender (TEY) or Triladyl. Those diluted sperms were then packed either in pool or straw and preserved at 5⁰C refrigerator. Sperm motility, viability and membrane integrity of each group were evaluated every 24 h for up to 5 days. Results showed that sperm motility in Triladyl of pool packaging system up to 3 days was higher than straw packaging system or TEY in pool or straw packaging system which were 45.8%, 26.1%, 32.1% and 9.1%, respectively (P<0.05). Percentage of sperm membrane integrity showed the same pattern to Triladyl both in pool and straw packaging system which was higher than TEY group (75.2% and 77,2%; P<0.05). Sperm viability in Triladyl both in pool or straw packaging system decreased (P<0.05) after 3 days of preservation (77.1% and 76.2%) but TEY significanly decreased after 4 days of preservation either in pool or straw packaging system (73.2% and 58.0%; P<0.05). It was concluded that sperm quality decreased with increasing of the length of preservation while Triladyl extender in pool packaging system showed the best quality.</p><strong>Key Words: </strong>Chilled Semen, Boer, Triladyl, Tris Egg Yolk, Straw

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Kriopreservasi Semen Kambing Boer dengan Konsentrasi Pengencer Nira Aren dan Gliserol Berbeda

Jurnal Ilmu dan Teknologi Peternakan Tropis ◽

10.33772/jitro.v6i1.5422 ◽

2019 ◽

Vol 6 (1) ◽

pp. 1

Author(s):

Muhammad Riyadhi ◽

Anis Wahdi ◽

Muhammad Rizal

Keyword(s):

Sperm Motility ◽

Membrane Integrity ◽

Egg Yolk ◽

Boer Goat ◽

Palm Juice ◽

Sperm Membrane ◽

Live Sperm

ABSTRAK Penelitian bertujuan untuk mengetahui efektivitas nira aren sebagai pengencer alternatif dalam proses pembekuan (kriopreservasi) semen kambing boer.Kriopreservasi semen kambing boer menggunakan pengencer tris-gliserol-kuning telur (P1 73-7-20%), nira aren-gliseol-kuning telur(masing-masing P2 74-6-20%, P3 73-7-20%, dan P4 72-8-20%) dan andromed (P5 tanpa mengandung kuning telur dan gliserol). Parameter evaluasi meliputi motilitas, viabilitas, dan membrane plasma utuh setelah pengenceran, ekuilibrasi dan thawing. Evaluasi motilitas pasca thawing menunjukkan P5 52% berbeda nyata (P<0.05) dengan P1 42%, selanjutnya P5 dan P1 berbeda sangat nyata (P<0.05) dengan P2 8%, P3 6% dan P4 12%. Viabilitas pasca thawing menunjukkan P5 65,4% tidak berbeda nyata (P>0,05) dengan P1 61,8%, akan tetapi P5 dan P1 berbeda sangat nyata (P<0.05) dengan P2 26,2%, P3 29,8%, dan P4 34%. Membran plasma utuh (MPU) pasca thawing menunjukkan P5 66,2% tidak berbeda nyata (P>0,05) dengan P1 65,4%, akan tetapi keduanya berbeda sangat nyata (P<0.05) dengan P2 39%, P3 38%, dan P4 36,2%. Disimpulkan kriopreservasi semen kambing boer dengan pengencer nira aren dan gliserol pada konsentrasi berbeda belum dapat dipergunakan sebagai sumber bibit berdasarkan standar nasional Indonesia.Kata Kunci : Kambing boer, semen, nira arenABSTRACTThe experiment was conducted to determine the effect of sugar palm juice as alternative extender for cryopreservation process of boer semen.Tris-glycerol-egg yolk (P1 73-7-20%), Sugar palm juice-glyserol-egg yolk (P2 74-6-20%, P373-7-20%, dan P4 72-8-20%), and andromed (P5) used as a extender in the cryopreservation process of boer semen. Sperm motility (%), live sperm (%) and sperm membrane integrity (%) were recorded after diluted, equilibration and freeze-thawing. Result of post thawing motility showed that P5 52% was significantly different (P <0.05) with P1 42%, then P5 and P1 were significantly different (P <0.05) with P2 8%, P3 6% and P4 12%. Viability after thawing showed P5 65.4% was not significantly different (P> 0.05) with P1 61.8%, but P5 and P1 significantly different (P <0.05) with P2 26.2%, P3 29.8 %, and P4 34%. Spermmembrane integrity post-thawing showed P5 66.2% was not significantly different (P> 0.05) with P1 65.4%, but both were very significantly different (P <0.05) with P2 39%, P3 38% and P4 36.2%. Conclusions, sugar palm juice-glycerol-egg yolk with differentconcentrationsineffectively as an alternative extenderin cryopreservation of boer semen.Keywords: boer goat, semen, sugar palm juice

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Macro- and microelements in serum and seminal plasma as biomarkers for bull sperm cryotolerance

Acta Veterinaria Scandinavica ◽

10.1186/s13028-021-00590-2 ◽

2021 ◽

Vol 63 (1) ◽

Author(s):

Maja Zakošek Pipan ◽

Petra Zrimšek ◽

Breda Jakovac Strajn ◽

Katarina Pavšič Vrtač ◽

Tanja Knific ◽

...

Keyword(s):

Seminal Plasma ◽

Semen Quality ◽

Sperm Quality ◽

Membrane Integrity ◽

Quality Characteristics ◽

Freezing And Thawing ◽

Additional Tool ◽

Bull Semen ◽

Bull Sperm ◽

Fresh Semen

ABSTRACT Background Wide variation in fertility rates is observed when using frozen bull semen, even when the bulls have met quality standards for semen production. Therefore, a simple and reliable test to assess the freezing potential of bull semen based on the analysis of fresh semen or blood would be of great value. Attention is now turning to assessment of seminal plasma components such as proteins and elements. In the present study, the concentrations of macro- and microelements in fresh bull semen plasma and in serum and their correlation with quality characteristics of fresh semen and with semen quality after freezing and thawing were determined. Ejaculates were collected from 30 mature bulls, and semen volume, concentration, sperm motility, morphology, tail membrane integrity, plasma membrane permeability and DNA fragmentation were determined on the day of collection and after freezing and thawing. The concentrations of macroelements (Na, Mg, K and Ca) and microelements (Cu, Fe, Zn and Se) were determined in the seminal plasma and serum. The semen samples were classified into satisfactory and unsatisfactory groups according to the fresh semen quality. Results Zinc and Se levels measured in serum were associated with almost all fresh and frozen-thawed semen quality characteristics, while Fe levels were associated only with acrosomal defects in fresh semen. Zinc and Fe levels in fresh seminal plasma were associated with various quality characteristics of fresh and frozen-thawed semen, while Se level in fresh seminal plasma was not associated with any of the semen quality characteristics. Conclusions Microelements were shown to be useful as biomarkers involved in the analysis of bull sperm quality and could be used as an additional tool to predict bull semen quality after freezing and thawing. Our results confirm that the analysis of Zn and Se levels in serum and Zn, Cu and Fe levels in fresh seminal plasma can provide information to discriminate between bull semen samples with spermatozoa with high or low cryotolerance.

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Sperm membrane integrity and stability after selection of cryopreserved ovine semen on colloidal solutions

Andrologia ◽

10.1111/and.12867 ◽

2017 ◽

Vol 50 (2) ◽

pp. e12867

Author(s):

T. G. Bergstein-Galan ◽

L. C. Bicudo ◽

L. Rodello ◽

R. R. Weiss ◽

S. D. Bicudo

Keyword(s):

Membrane Integrity ◽

Colloidal Solutions ◽

Sperm Membrane ◽

Selection Of

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Seminal plasma proteins protect flow-sorted ram spermatozoa from freeze - thaw damage

Reproduction Fertility and Development ◽

10.1071/rd08238 ◽

2009 ◽

Vol 21 (4) ◽

pp. 571 ◽

Cited By ~ 23

Author(s):

T. Leahy ◽

J. I. Marti ◽

G. Evans ◽

W. M. C. Maxwell

Keyword(s):

Seminal Plasma ◽

High Speed ◽

Sperm Quality ◽

Membrane Integrity ◽

Weight Fraction ◽

Protein Supplementation ◽

Freezing Process ◽

Spermatozoa Motility ◽

Functional Integrity

Seminal plasma improves the functional integrity of compromised ram spermatozoa but has been reported to be toxic to sorted spermatozoa. The present study attempted to clarify this paradoxical effect and improve the functional integrity of spermatozoa following sorting and cryopreservation. The in vitro function of sorted spermatozoa (motility characteristics and membrane integrity) was examined after supplementation with differing concentrations and protein fractions of seminal plasma at various stages of the sorting and freezing process. For all experiments, spermatozoa (two males, n = four ejaculates per male) were processed through a high-speed flow cytometer before cryopreservation, thawing and incubation for 6 h (37°C). Supplementation of crude seminal plasma (CP), its low molecular weight fraction (LP; <10 kDa) or protein-rich fraction (SPP; >10 kDa), immediately before freezing improved the functional integrity of sorted spermatozoa compared with no supplementation (control), whereas supplementation after thawing had no effect for CP and LP. The protective effect of seminal plasma was not altered by increasing the amount of protein supplementation. No toxic effect of CP, SPP or LP was evident even when supplemented at high protein concentrations. It is concluded that seminal plasma protein, if added to ram spermatozoa after sorting and before freezing, can improve post-thaw sperm quality and consequently the efficiency of sorting. This effect is most likely related to protection of the spermatozoa during freeze–thawing.

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