Effect of cooling rates on post-thaw sperm motility, membrane integrity, capacitation status and fertility of dairy bull semen used for artificial insemination in sweden

1999 ◽  
Vol 52 (4) ◽  
pp. 641-658 ◽  
Author(s):  
A. Januskauskas ◽  
J. Gil ◽  
L. Söderquist ◽  
M.G.M. Hrd ◽  
M.Ch. Hrd ◽  
...  
2017 ◽  
Vol 29 (3) ◽  
pp. 490 ◽  
Author(s):  
Asmatullah Kaka ◽  
Wahid Haron ◽  
Rosnina Yusoff ◽  
Nurhusien Yimer ◽  
A. M. Khumran ◽  
...  

This study was conducted to investigate the effect of docosahexanoic acid (DHA) supplementation in BioXcell extender on the quality of frozen–thawed bull semen. Twenty-four ejaculates were collected from three bulls (eight from each bull). Ejaculates with motility ≥70% and normal morphology ≥80% were extended into BioXcell extender to which 0 (control), 3, 5, 10 or 15 ng mL–1 DHA was added. The supplemented semen samples were incubated at 37°C for 15 min for DHA uptake by spermatozoa. Later, samples were cooled for 2 h at 5°C and packaged into 0.25-mL straws, frozen in liquid nitrogen for 24 h and subsequently thawed for evaluation. Results are presented as percentages ± s.e.m. Supplementation with DHA at 3 ng mL–1 significantly improved sperm functional parameters including sperm motility, normal morphology, viability, acrosome integrity and membrane integrity when compared with other supplemented groups and the control. Lipid peroxidation increased as the incorporation of DHA supplementation increased. In conclusion, 3 ng mL–1 concentration of DHA resulted in superior quality of frozen–thawed bull spermatozoa and is suggested as the optimum level of DHA to be added into BioXcell extender.


2020 ◽  
Vol 53 (1) ◽  
Author(s):  
Bushra Ismail Khan ◽  
Shamim Akhter ◽  
Sanwal Aslam ◽  
Rabea Ejaz

The current study was planned to evaluate the supplementation of Polyvinylpyrrolidone in extender on cryopreservation of Nili-Ravi buffalo bull semen. The semen samples were collected from Nili-Ravi buffalo (Bubalus bubalis) bull kept at SPU Qadirabad, District Sahiwal, Pakistan. Qualifying semen ejaculates having motility >60%, volume >5-6ml and concentration >0.5 billion/ ml were diluted 50 × 106 motile sperm ml approximately at 37°C in Tris-citric acid extender supplemented with different concentrations of PVP (0.01, 0.05, 0.1mM). The extender without PVP was kept as control. Semen was stored at 4°C for a period of 2 h and kept at 4°C for 4h. Semen was filled in 0.5 ml French straws using suction pump at 4°C, plunged and stored in liquid nitrogen (-196°C). Semen straws were rewarmed at 37°C for 30 seconds and assessed for sperm motility, plasma membrane integrity (PMI), dead sperm percentage and the live sperm percentage. The data on the role of PVP on different parameters of semen quality were analyzed by using ANOVA and RCBD. Higher percentage (P< 0.05) of sperm motility (66.1±7.51 and 59.4±10.72) and PMI (72.9±5.39 and 75.7±6.5) was observed in extenders having 0.05 mM and 0.1mM PVP compared to extenders having 1.5mM PVP and control. The percentage acrosomal integrity was observed greater (P< 0.05) in extended semen containing 0.1mM (68.2±0.50) PVP compared to extenders having 0.01 and control.


2019 ◽  
Vol 19 (3) ◽  
pp. 733-746
Author(s):  
Marek Lecewicz ◽  
Rafał Strzeżek ◽  
Anna M. Majewska ◽  
Piotr S. Purpurowicz ◽  
Władysław Kordan

AbstractArtificial insemination (AI) and semen cryopreservation are the most accessible and commonly used techniques for breeding domestic animals. Among many parameters, such as plasma membrane integrity and acrosome structure, one of the key factors that determine the quality of frozen-thawed samples for artificial insemination is sperm motility. Sperm motility is one of the key parameters that determine the quality of frozen-thawed samples for AI. The total number of progressively motile spermatozoa in thawed canine semen is correlated with fertility. A variety of substances were used to compare sperm motility with the control. The aim of this study was to determine the effect of semen extender supplementation with motility stimulants, pentoxifylline (PTX), caffeine (CAF) and 2’-deoxyadenosine (DX), after different post-thaw incubation times (30, 60, 120 min) on the motility, selected kinematic parameters, plasma membrane integrity and mitochondrial membrane potential of cryopreserved canine spermatozoa. During attempts to improve the quality of cryopreserved semen, the applied substances exerted beneficial effects at a concentration of 10 mM. We demonstrated that both phosphodiesterase inhibitors, caffeine and pentoxifylline, as well as 2’-deoxyadenosine increased the motility and selected kinematic parameters of thawed canine spermatozoa.


2015 ◽  
Vol 27 (5) ◽  
pp. 784 ◽  
Author(s):  
Mushtaq Ahmad ◽  
Nasim Ahmad ◽  
Amjad Riaz ◽  
Muhammad Anzar

This study was designed to compare the kinetics of sperm survival in different types of bull semen. Fresh ejaculates from four bulls were pooled, diluted in Tris-citric acid-egg yolk-glycerol extender, cooled to 4°C, frozen in LN2 and thawed at 37°C. Fresh, diluted, cooled and frozen–thawed semen were incubated at 37°C, and evaluated at 0, 2, 4, 6, 12 and 24 h after the beginning of incubation. In Experiment 1, progressive sperm motility, normal acrosomes and plasma membrane integrity and asymmetry were determined. In Experiment 2, generation of superoxide anion (O2•) along with plasma membrane permeability and generation of hydrogen peroxide (H2O2) along with plasma membrane integrity were assessed. In Experiment 1, frozen–thawed semen had shorter survival times for progressive sperm motility, and spermatozoa with intact plasma membranes and acrosomes (IPM-IACR) as compared with other types of semen (P < 0.05). Fresh spermatozoa underwent a necrotic pathway, diluted and cooled spermatozoa underwent an apoptosis-like pathway and frozen–thawed spermatozoa underwent both necrotic and apoptosis-like pathways. In Experiment 2, spermatozoa in all four types of semen exhibited O2•– generation and increased plasma membrane permeability, and became necrotic without H2O2 generation during incubation (P < 0.05). In conclusion, frozen–thawed semen had shorter sperm longevity, which has important implications relating to the timing of artificial insemination. Different types of semen followed different death pathways. During incubation, spermatozoa in all types of semen generated O2•–, which increased the permeability and compromised the integrity of the plasma membrane.


2014 ◽  
Vol 26 (1) ◽  
pp. 140
Author(s):  
M. M. Seshoka ◽  
M. L. Mphaphathi ◽  
T. L. Nedambale

Cryopreservation has been reported to damage approximately 40 to 50% of viable spermatozoa in bulls. It is critical to evaluate frozen-thawed spermatozoa with computer-aided sperm analysis (CASA) and find a suitable cryoprotectant for Nguni semen. The study was conducted to compare cryo-effectiveness of glycerol (GLY), dimethyl sulfoxide (DMSO), propanediol (PND), and ethylene glycol (EG) cryoprotectants at 12% concentrations during freezing of Nguni bull semen. Semen was collected from 18 stud Nguni bulls of proven fertility with the use of an electro-ejaculator. Collected semen samples were kept in a thermo flask (37°C) and transported to the laboratories. Semen samples were pooled and sperm motility rate was evaluated using CASA. Semen was then diluted (1 : 2; vol : vol) with egg-yolk citrate extender supplemented with either 12% GLY, DMSO, PND, or EG. Semen samples were equilibrated for 4 h at 5°C. After equilibration, samples were loaded into 0.25-mL straws and placed into the controlled rate programmable freezer using a customized freezing curve (from 5 to –5°C at 0.008°C min–1 and from –3 to – 130°C at 6°C min–1). Following thawing of semen, artificial insemination was conducted on 104 oestrus-synchronised Nguni cows. The IVF was also conducted on 120 oocytes to check the cleavage rate. Data were analysed using ANOVA. There was a significant difference (P < 0.05) between raw total sperm motility (94.70 ± 2.63) and frozen-thawed sperm total motility with GLY (77.80 ± 11.03), EG (20.35 ± 11.86), DMSO (15.68 ± 10.14), and PND (11.19 ± 11.27) groups. The pregnancy rate following artificial insemination was 75.9% and a total of 86.6% oocytes had cleaved after fertilization with frozen (12% GLY)/thawed semen. In conclusion, cryopreservation process reduced sperm motility and velocity rates, regardless of cryoprotectant. Egg-yolk citrate extender supplemented with 12% glycerol had recorded the highest post-thaw sperm motility rate.


2016 ◽  
Vol 19 (1) ◽  
pp. 147-158 ◽  
Author(s):  
M. Lecewicz ◽  
W. Kordan ◽  
A. Majewska ◽  
S. Kamiński ◽  
A. Dziekońska ◽  
...  

Abstract The aim of the study was to determine the effects of platelet-activating factor (PAF) on selected quality parameters of cryopreserved bull semen with reduced sperm motility used for artificial insemination. The aim of experiment 1 was to identify the optimal concentration of the phospholipid able to preserve sperm viability. Cryopreserved semen was treated with different PAF concentrations: 1×10-5M, 1×10-6M, 1×10-7M, 1×10-8M and 1×10-9M. The experiment demonstrated that PAF at concentration 1×10-9M increased most the sperm viability parameters (motility parameters, plasma membrane integrity and mitochondrial function) after 120 min of incubation of thawed semen at 37°C. Cryopreserved bull semen with reduced sperm motility (below 70%) was supplemented with PAF in a concentration of 1×10-9M. A statistically significant increase in sperm motility, percentage of linear motile spermatozoa and VSL value was observed after 120 min incubation of sperm with 1×10-9M PAF. Sperm supplementation with PAF also had positive effects on plasma membrane integrity and percentage of spermatozoa with preserved mitochondrial transmembrane potential, but the differences were not statistically significant. The results indicated positive effects of PAF supplementation at a concentration of 1×10-9M on the selected sperm quality parameters in cryopreserved bull semen with reduced motility.


2008 ◽  
Vol 20 (1) ◽  
pp. 128 ◽  
Author(s):  
B. Szczesniak-Fabianczyk ◽  
M. Bochenek ◽  
A. T. Palasz ◽  
J. De la Fuente ◽  
Z. Smorag

Replacement of animal-origin components in extenders used for bull semen freezing is of high importance for individuals involved in cattle breeding. The experiment was designed to compare efficacy of 5 different semen extenders in cryopreservation of bull semen: sodium citrate-based extender containing egg yolk (CT), commercially available Bioxcell� (IMV Technologies, L'Aigle, France), and 3 custom-made homogenized plant lipidsbased, egg yolk-free extenders (Y-1, Y-2, and Lipo) . The objective was to determine whether homogenization procedures of lipids improve the quality of the extender. Lipid homogenates of custom-made extenders were prepared in Tris buffer using a high pressure homogenizer (Nira Saovi, Parma, Italy). Ten (Y-1) or 5 (Y-2) homogenization cycles were applied and then 8% glycerol was added. Lipid liposomes were produced by simultanous high pressure homogenization of lipids and glycerol supplementation (Lipo). Semen was collected from young bulls of 3 different breeds (Simmental, Polish Red, and Holstein; 1 ejaculate/bull). Each ejaculate with at least 70% motility was split into 5 parts and processed further by a standard freezing protocol: semen was diluted at 35�C with each of the 5 extenders to a concentration of 100 � 106 spermatozoa per mL, cooled to 4�C over 5 h, aspirated into 0.25-mL plastic straws, frozen in liquid nitrogen vapor to –140�C, and then plunged into LN2. Straws were thawed in a water bath at 37�C for 20 s. Sperm motility was estimated microscopically immediately after thawing and after 5 h of storage at 22�C. Immediately after thawing, flow cytometry and SYBR-14/PI staining were used for examination of sperm membrane integrity (live/dead assay). A total of 20 000 spermatozoa of each sample were counted. Student's t-test was used to estimate statistical differences between experimental groups. The mean sperm motility after thawing ranged from 45.6% (SD = 13.7) for CT (egg yolk extender) to 57.8% (SD = 7.1) for Lipo. A significant difference (P < 0.05) was observed betweenY-1 (50.0%, SD = 9.7) and Lipo and Bioxcell (56.1%, SD = 8.6). After 5 h of storage at 22�C, the mean motility for all tested bulls ranged from 25.0% (SD = 7.1) for CT to 42.2% (SD = 7.5) for Lipo. Significant differences were observed between Lipo (P < 0.01), Y-2 (P < 0.05) and CT, and between Y-1 and Lipo (P < 0.01). Mean percentage of 'live' spermatozoa with intact membrane after freezing/thawing was 51.85% (SD = 11.49) for Y-1, 45.72% (SD = 9.36) for Y-2, 47.57% (SD = 7.93) for Lipo, 45.47% (SD = 8.35) for Bioxcell, and 49.06 (SD = 11.59) for CT. No significant differences were observed except forY-1 and Bioxcell extenders (P < 0.05). It can be concluded that both methods of lipid/glycerol homogenization can be successfully applied in the preparation of bull semen extender. In addition, extensive lipid homogenization (10 cycles) produced more transparent extender that in turn improved visualization of sperm. Custom-made plant origin lipids homogenization may provide a valuable alternative for the preparation of extenders that more closely match the membrane composition of bull sperm cells and thus contribute to development of an efficient extender free of animal-origin components for bull semen freezing.


Moringa oleifera extract is a strong natural antioxidant that when was added to the semen extenders, it induced a cryoprotection to spermatozoa effect through elimination of the excess free radicals. So, the existing study intended for clarification of the consequence of extract of Moringa leaves (MLE) on bull spermatozoa after chilling and cryopreservation. MLE concentrations were 0% (control), 10, 20, 30, 40 and 50% (v/v) [MLE: TCF (Tris-citric-fructose diluent)] then 20% egg yolk was added, then the extended semen was assigned to the freezing protocol. Then, it was evaluated for (motility, alive, abnormality %, sperm membrane integrity % before and after freezing). Sperm motility was kept high with the concentration 10, 30 and 40% of MEEY till 8 days of chilling. The concentration 20% maintained sperm motility high till 7 days of chilling. Addition of MLE to TCF significantly (P<0.002) improved sperm motility in all concentrations except the 50% moringa enriched extender with egg yolk (MEEY) where sperm motility was maintained as the control. The use of MEEY maintained % of alive sperms and % of normal spermatozoal membrane (HOST%) as good as the control. In conclusion: moringa as a herbal supplement to semen diluents enhanced preservation in cooled and cryopreserved cattle bull semen.


2019 ◽  
Vol 1 (1) ◽  
pp. 22-26
Author(s):  
Jamal Mehmood shah ◽  
Tahir Hameed ◽  
Farhat abbas Bokhari ◽  
Gul zaman

TThe   study   was  carried   out   during the yearr  2017   to   determine   the   fresh   and   post-thaw   quality   of   four   Holstein   Friesian   bulls.   Semen   samples   were   examined   for ejaculate   volume,   pH,   mass   activity,   sperm   concentration   and   sperm   motility and   sperm   membrane   integrity   (HOST).   The   ejaculated   semen   volume   of   Holstein   Friesian   bulls   was   in   the   range   of   6-8ml   and   differences   in   ejaculates   were   significant   (P<0.01).   The   volume   was   significantly   (P<0.05)   higher   (7.75ml)   when   collected   on   5th   July,   slightly   decrease   in   volume   (7.25ml)   when   collected   on   12th   May   and   26th   July.   Semen   pH   was   higher   (6.65)   for   26th   July   ejaculation   and   lowest (6.55ml)   for   24th   May   ejaculation.   The   results   indicated   that   the   semen   of   Holstein   Friesian   bulls   did   not   have   considerable   variation   in   pH   during   May   –   July.   Most   of   the   semen   samples   were   creamy   white and   yellow   in   colour,   while   few   samples   were   milky   white   and   watery.   Mass   activity   score   of   semen   samples   indicated   vigorous   movement with   moderate rapid   waves   and   eddies.   The   sperm   motility   of   fresh   vs   post-thaw   semen   was   obtained   72.80   vs   48.75   percent   (12th   May),   75.00   vs   48.75   percent   (24th   May),   76.25   vs   55.00   percent   (2nd   June),   73.75   vs   52.50   percent   (21st   June),   75.00   vs   56.25   (5th   July)   and   75.00   vs   50.00   percent   (26th   July).   In   case   of   post-thaw   semen,   highest   sperm   motility   of   56.25   percent   was   recorded   in   semen   ejaculated   on   5th   July,   while   lowest   post-thaw   sperm   motility   of   48.75   percent   was   observed   in   semen   ejaculated   on   12th   May   and   24th   May.   Highest   sperm   concentration   of   1549.75x106   was   determined   in   semen   ejaculated   on   21.06.2017   and   lowest   concentration   (1259.50x106)   in   26.07.2017   collected   samples.   It   was   concluded   that   sperm   motility   was   significantly   (P<0.01)   affected   by   semen   types   (fresh   and   post-thaw),   while   sperm   concentration   was   also   significantly   (P<0.01)   affected   by   ejaculation   date   and   bulls.   The   membrane   integrity   (HOST)   of   the   pooled   data   on   fresh   semen   samples   over   a   period   of   three   months   was   58.37   percent   against   post-thaw   membrane   integrity   of   44.79   percent.


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