Effect of docosahexanoic acid on quality of frozen–thawed bull semen in BioXcell extender

2017 ◽  
Vol 29 (3) ◽  
pp. 490 ◽  
Author(s):  
Asmatullah Kaka ◽  
Wahid Haron ◽  
Rosnina Yusoff ◽  
Nurhusien Yimer ◽  
A. M. Khumran ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Membrane Integrity ◽  
Optimum Level ◽  
Docosahexanoic Acid ◽  
Normal Morphology ◽  
Bull Semen ◽  
Acrosome Integrity

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.

scholarly journals Comparative effects of addition of superoxide dismutase and reduced glutathione on cryopreservation of Sahiwal bull semen

2019 ◽  
Vol 69 (4) ◽  
pp. 1291
Author(s):  
A. Murtaza ◽  
M. Ahmad ◽  
M. Zubair ◽  
S. Umar ◽  
A. Mushtaq ◽  
...  
Keyword(s):  
Superoxide Dismutase ◽  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Semen Quality ◽  
Reduced Glutathione ◽  
Membrane Integrity ◽  
Bull Semen ◽  
Microscopic Evaluation ◽  
Artificial Vagina

The present study aimed to investigate effects of superoxide dismutase (SOD) and reduced glutathione (GSH) on the quality of frozen-thawed semen of Sahiwal bulls. Semen was collected twice a week for 8 weeks by artificial vagina from six Sahiwal bulls, kept at the Semen Production Unit Qadirabad, Sahiwal-Pakistan. After gross and microscopic evaluation, qualifying semen ejaculates were divided into 10 equal aliquots and diluted in extenders enriched with no antioxidants (control); or supplemented with either SOD (50, 100 and 200 IU/mL), or GSH (0.5, 1 and 2 mM) or their combinations (50 IU/mL SOD and 0.5 mM GSH, 100 IU/mL SOD and 1 mM GSH and 200 IU/mL SOD and 2 mM GSH). Samples were then frozen and stored in liquid nitrogen at -196°C for 24 h. The following parameters were evaluated for semen quality: post-thawed sperm motility, viability, acrosome and membrane integrity. According to the results, sperm motility, viability, acrosome and membrane integrity were significantly (P<0.05) higher in samples treated either with 100 IU/mL of SOD; 1 mM and 2 mM of GSH or 50 IU/mL of SOD plus 0.5 mM of GSH. In conclusion, semen quality might be improved by supplementing semen extenders with 100 IU/mL of SOD; 0.5 and 1 mM of GSH and combination of 50 IU/mL and 0.5 mM of SOD and GSH, respectively.

scholarly journals Spectrophotometric Analysis of the Resazurin Reduction Test as a Tool for Assessing Canine Semen Quality

2013 ◽  
Vol 57 (2) ◽  
pp. 281-285 ◽  
Author(s):  
Rafał Strzeżek ◽  
Krystyna Filipowicz ◽  
Marta Stańczak ◽  
Władysław Kordan
Keyword(s):  
Sperm Motility ◽  
Seminal Plasma ◽  
Semen Quality ◽  
Membrane Integrity ◽  
Spectrophotometric Analysis ◽  
Normal Morphology ◽  
Additional Tool ◽  
Reduction Test ◽  
Highly Correlated

Abstract The resazurin reduction test (RRT) was subjected to spectrophotometric analysis to evaluate the quality of canine semen. Twenty four samples of canine semen were analysed. The absorption peaks for resazurin and resorufin were determined at 615 and 580 nm, respectively. The RRT ratio (RRTsperm-the ratio for samples containing spermatozoa, RRTplasma-the ratio for samples containing seminal plasma) was calculated by dividing the absorbance at 580 nm by the absorbance at 615 nm. Spearman’s correlation test was used to determine the significance of correlations between the analysed sperm parameters and the results of the resazurin reduction assay. The RRT ratio was highly correlated with sperm motility (r=0.68, P<0.01), progressive sperm motility (r=0.61, P<0.01), the subpopulation of cells with rapid velocity (r=0.72, P<0.01), and the subpopulation of cells with medium velocity (r= -0.54, P<0.05). A negative correlation was observed between the reducing capacity of seminal plasma vs. sperm with plasma membrane integrity (r= -0.60, P<0.01) and sperm with normal morphology (r= -0.58, P<0.01). The RRT test can be used as an additional tool for evaluation of the quality of canine semen.

scholarly journals Effects of the addition of docosahexaenoic acid and ?-tocopherol on quality of equine spermatozoa stored at 5°C

2020 ◽  
Vol 41 (1) ◽  
pp. 167 ◽  
Author(s):  
Breno Fernandes Barreto Sampaio ◽  
Bruno Gomes Nogueira ◽  
Maria Inês Lenz Souza ◽  
Eliane Vianna da Costa-e-Silva ◽  
Carmem Estefânia Serra Neto Zúccari
Keyword(s):  
Lipid Peroxidation ◽  
Plasma Membrane ◽  
Docosahexaenoic Acid ◽  
Sperm Motility ◽  
Membrane Integrity ◽  
Control Group ◽  
Mitochondrial Activity ◽  
Membrane Composition ◽  
Artificial Vagina

Plasma membrane composition has impact on phase transition from liquid crystal to gel state of cooled sperm cell. The incorporation of polyunsaturated fatty acids increases its fluidity and can contribute to sperm motility. The aim of this study was to compare the effect of adding docosahexaenoic acid (DHA) and ?-tocopherol (?-Toh) to the cooling extender, singly or combined, to the equine sperm parameters, submitted to cooling, up to 72 hours. Two ejaculates of ten stallions collected with artificial vagina were used, and evaluated for motility, plasma membrane integrity, chromatin fragmentation, mitochondrial activity and lipid peroxidation, according to the following treatments: C; DHA; ?-Toh; DHA/?-Toh; EtOH 100: and EtOH 140 (corresponding to control; 10 ng mL-1 of DHA; 2 mM of ?-Toh; : 10 ng mL-1 of DHA + 2 mM of ?-Toh; 100 µL of ethanol and 140 µL of ethanol respectively). DHA treatment showed higher motility (68.2 ± 12.3; p < 0.05) when compared to control (62.1 ± 16.2), DHA/?-Toh (61.3 ± 12.7) and EtOH (58.1 ± 8.6) groups. In lipid peroxidation assay, the control group showed 2,506.2 ± 796.4 ng of MDA 108 spermatozoa-1, being significantly higher (p < 0.05) than the groups treated with DHA (2,036.0 ± 687.0), ?-Toh (1,890.8 ± 749.5) and DHA/?-Toh (1,821.1 ± 627.2). In conclusion, ?-Toh was effective in diminishing lipid peroxidation of equine sperm subjected to cooling, and DHA improved sperm motility and, in spite of being a polyunsaturated fatty acid with high susceptibility to peroxidation, reduced lipid peroxidation.

Evaluate the fresh and Post Thawing Quality of Holstein Friesian Bull semen

2019 ◽  
Vol 1 (1) ◽  
pp. 22-26
Author(s):  
Jamal Mehmood shah ◽  
Tahir Hameed ◽  
Farhat abbas Bokhari ◽  
Gul zaman
Keyword(s):  
Sperm Motility ◽  
Sperm Concentration ◽  
Membrane Integrity ◽  
Pooled Data ◽  
Bull Semen ◽  
Holstein Friesian ◽  
Semen Volume ◽  
Sperm Membrane ◽  
Mass Activity

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.

scholarly journals Ram and Goat Semen Immunosexed and Diluted in Powdered Coconut Water-Based Preservation Medium (ACP101/102c).

2021 ◽  
Vol 49 ◽  
Author(s):  
Bruna Farias Brito ◽  
Bárbara Mara Bandeira Santos ◽  
Leonardo Alves Rodrigues Cabral ◽  
Luiz Carlos Pinheiro Maia ◽  
Natanael Aguiar Braga Negreiros ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Specific Protein ◽  
Coconut Water ◽  
In Vivo Studies

Background: Sperm sexing aims to separate sperm populations in carriers of the “X” or “Y” chromosome. Currently, flow cytometry is a technique that allows greater accuracy; however, it causes structural changes in sperm, reduces viability, and has a high cost. As a result, other methods have been researched, including immunosexing, which uses monoclonal antibodies to detect sex-specific surface antigens. Thus, the objective of this study was to evaluate the immunosexing technique using a monoclonal antibody against sex-specific protein (HY) in the conservation of ram and goat semen in ACP101/102c.Materials, Methods & Results: Ejaculates from 5 rams and 5 goats were collected with the aid of an artificial vagina; they were evaluated and submitted to the immunosexing protocol, according to the manufacturer's recommendations, using the Monoclonal Antibody Kit specific for mammalian sperm with “Y” chromosomes (HY; HY Biotechnology, Rio de Janeiro, RJ, Brazil). After sexing, the supernatant was resuspended in the cryopreservation diluent: ACP ram (ACP101/102c + 20% egg yolk + 7% glycerol) and ACP goat (ACP101/102c + 2.5% egg yolk + 7% glycerol), packaged in 0.25 mL straws, refrigerated at 4°C, stabilized for 30 min, frozen in liquid nitrogen vapor (-60°C) for 15 min, immersed in liquid nitrogen, and stored in cryogenic cylinders. The samples were evaluated in natura (T1), after immunosexing (T2) and after thawing (T3) for sperm motility subjectively using conventional microscopy (40x). Plasma membrane integrity (IMP) and sperm cell morphology were evaluated by the smear staining technique using eosin-nigrosine dye, and the percentages of healthy and morphologically defect spermatozoa were determined. In the evaluation of ram semen regarding sperm motility and IMP, no statistically significant differences were observed between treatments after sexing in the evaluation of absolute data (P > 0.05), with the difference being observed only between T1 and T2, and T3 (P < 0.05). Regarding the relative percentage and sperm morphology, no statistically significant differences were observed (P > 0.05). Regarding the evaluation of goat semen samples, the motility parameters were consistent with the technique submitted; however, the IMP data did not appear as expected, requiring further evaluation for a better assessment of the technique for this species.Discussion: The data obtained from ram semen submitted to the immunosexing protocol, regarding the absolute evaluation of motility and IMP, demonstrated that the non-sexed semen (T1) was superior to the sexed treatments (T2 and T3); however, it is noteworthy that freezing started with approximately 50% of the cells, since the immunosexing technique results in a loss of viability of approximately 50% of the sperm, which corresponds to the ratio of sperm carrying the X chromosome. In addition, when the data in this study were transformed into relative values, no statistical differences were observed, indicating that the immunosexing protocol, as well as the freezing protocol, did not significantly affect the quality of ram sperm cells. In relation to the immunosexing of goat semen, future studies should be conducted in vitro to define a more appropriate protocol for the species and, in addition, in vivo studies should be performed to prove the quality of the technique. It was concluded that the immunosexing process using a monoclonal antibody against sex-specific protein (HY) associated with the use of powdered coconut water diluent (ACP101/102c) in the cryopreservation of semen proved to be efficient in the in vitro evaluation of ovine species.

Quality of ram spermatozoa separated with modified swim up method

2018 ◽  
Vol 33 (2) ◽  
pp. 62-70 ◽  
Author(s):  
A Hossain ◽  
MM Islam ◽  
F Naznin ◽  
RN Ferdousi ◽  
FY Bari ◽  
...  
Keyword(s):  
Semen Quality ◽  
Membrane Integrity ◽  
Quality Parameters ◽  
Fluid Medium ◽  
Normal Morphology ◽  
The Mean ◽  
Mass Activity ◽  
Fresh Semen ◽  
Artificial Vagina

Semen was collected from four rams, using artificial vagina and viability%, motility% and plasma membrane integrity% were measured. Fresh ejaculates (n = 32) were separated by modified swim-up separation using modified human tubal fluid medium. Four fractions of supernatant were collected at 15-minute intervals. The mean volume, mass activity, concentration, motility%, viability%, normal morphology and membrane integrity% (HOST +ve) of fresh semen were 1.0 ± 0.14, 4.1 ± 0.1 × 109 spermatozoa/ml, 85.0 ± 1.3, 89.4 ± 1.0, 85.5 ± 0.7, 84.7 ± 0.5 respectively. There was no significant (P>0.05) difference in fresh semen quality parameters between rams. The motility%, viability% and HOST +ve % of first, second, third and fourth fractions were 53.4 ± 0.5, 68.2 ± 0.3, 74.8 ± 0.3 and 65.5 ± 0.4; 55.5 ± 0.4, 66.2 ± 0.4, 74.5 ± 0.3 and 73.6 ± 0.3 and 66.7 ± 0.5, 66.8 ± 0.5, 65.2 ± 0.4 and 74.7 ± 0.5 respectively. The motility%, viability% and membrane integrity% of separated semen samples differed significantly (P<0.05) between four fractions. The mean motility% and viability% were significantly higher (P<0.05) in third fraction (74.8 ± 0.3%), whereas the mean HOST +ve% was significantly higher (P<0.05) in fourth fraction (74.7 ± 0.5). All quality parameters of separated spermatozoa were significantly (P<0.05) lower than that of fresh semen. The pregnancy rates were higher with fresh semen (71%) in comparison to that of separated sample (57%).Bangl. vet. 2016. Vol. 33, No. 2, 62-70

scholarly journals Cooling of equine semen at 16°C for 36 hours with addition of different glutathione concentrations

2015 ◽  
Vol 36 (6) ◽  
pp. 3699
Author(s):  
Rodrigo Arruda de Oliveira ◽  
Marco Antônio De Oliveira Viu ◽  
Maria Lúcia Gambarini
Keyword(s):  
Lipid Peroxidation ◽  
Sperm Motility ◽  
Membrane Integrity ◽  
Membrane Lipid ◽  
Sperm Cells ◽  
Sperm Viability ◽  
Membrane Lipid Peroxidation ◽  
Acrosomal Membrane ◽  
In Vitro Effects

Handling equine semen during the refrigeration process reduces sperm viability, and consequently causes membrane lipid peroxidation, among other challenges. The present study aimed to evaluate the in vitro effects of glutathione (control, 1. 0, 1. 5, and 2. 5 mM) on equine semen in a refrigeration protocol of 16ºC for 36 hours. The following variables were evaluated after 0, 12, 24, and 36 hours refrigeration: total sperm motility, vigor, viability, and plasma and acrosomal membrane integrity. Motility was higher with 2. 5mM of glutathione (57. 8 ± 7. 3) after 12 hours of refrigeration compared to the control (53. 2 ± 8. 3) (P < 0. 05). After 36 hours of refrigeration, motility was higher with 1. 5 mM (43. 4 ± 12. 7) and 2. 5mM glutathione (45. 5 ± 6. 2), than it was with 1mM glutathione (38. 2 ± 9) and the control (35. 5 ± 18. 4) (P < 0. 05), respectively. Vigor was highest with 1. 5mM glutathione (3. 7 ± 0. 3) after 36 hours compared to the control (3. 2 ± 1. 1), (P < 0. 05). Viability differed between control and 1mM treatments (79. 5 ± 1. 8) only after 24 hours (75. 5 ± 9. 7) (P < 0. 05). Throughout the investigation, no significant differences were noted in plasma and acrosomal membrane integrity (P > 0. 05). The 1. 5 and 2. 5mM glutathione levels were more efficient in protecting sperm cells and yielded higher total motility values after 36 hours of refrigeration.

scholarly journals Antioxidant effect of Mn2+ on capacitation and acrosome reaction of fresh and chilled cattle bull semen

2012 ◽  
Vol 1 (1) ◽  
pp. 18
Author(s):  
Amrit Kaur Bansal ◽  
Ranjna Sundhey Cheema ◽  
Vinod Kumar Gandotra
Keyword(s):  
Reactive Oxygen Species ◽  
Lipid Peroxidation ◽  
Acrosome Reaction ◽  
Reactive Oxygen ◽  
Antioxidant Effect ◽  
Oxygen Species ◽  
Sperm Capacitation ◽  
Bull Semen

The aim of this paper was to investigate the antioxidant effect of Mn2+ (200 mM) on the sperm capacitation and acrosome reaction of fresh and chilled cattle bull semen. It has been found that Mn2+ supplementation improves (P≤0.05) the motility at 0, 2, 4 and 6 h of incubation. MDA (malondialdehyde), end product of lipid peroxidation, decreases significantly (P≤0.05) with the supplementation of manganese at 0- and 6-hr of incubation both in fresh and chilled semen. Manganese also increases acrosome reaction significantly (P≤0.05) both in fresh and chilled semen at 0, 4 and 6 h of incubation. Therefore, our findings suggest the role of Mn2+supplementation in improving the quality of cattle bull semen by its scavenging property<em> i.e.</em> reduction in the production of reactive oxygen species during its storage at 4°C or incubation at 37°C for capacitation.

scholarly journals COMBINED EFFECT OF STREPTOMYCIN AND SWEET ORANGE ESSENTIAL OIL TO MEMBRANE AND ACROSOME INTEGRITY BOER GOAT FROZEN SEMEN

2019 ◽  
Vol 3 (2) ◽  
pp. 89
Author(s):  
Sukma Aditya Sitepu ◽  
Julia Marisa
Keyword(s):  
Essential Oil ◽  
Sweet Orange ◽  
Membrane Integrity ◽  
Boer Goat ◽  
Randomized Design ◽  
Frozen Semen ◽  
Acrosome Integrity ◽  
Completely Randomized Design ◽  
Orange Essential Oil

One of factors that cause a bad quality of Boer Goat frozen semen is the growth of bacterial. This can be overcome by adding antibiotics such as streptomycin. To further suppress the growth of bacteria can be added other ingredients that contain antibacterials such as sweet orange essential oil. The purpose of this research is to know the percentage value of Membrane Integrity and Acrosome Integrity on Boer Goat frozen semen with addition sweet orange essential oil and streptomycin. The method used was experimental using Completely Randomized Design with 5 treatments and 5 replications. The treatment in this research is addition 0%, 0,25%; 0.5%; 0.75% and 1% sweet orange essential oil on tris yolk and streptomycin extender. The results showed the best treatment addition combination streptomycin and sweet orange essential oil to percentage Membrane Integrity and Acrosome Integrity is increase 1% sweet orange essential oil.Keywords: Boer Goat, essential oil, frozen semen, streptomycin, sweet orange.

scholarly journals Effect of Supplementation of Polyvinylpyrrolidone in Extender on Buffalo Semen Parameters

2020 ◽  
Vol 53 (1) ◽  
Author(s):  
Bushra Ismail Khan ◽  
Shamim Akhter ◽  
Sanwal Aslam ◽  
Rabea Ejaz
Keyword(s):  
Sperm Motility ◽  
Semen Quality ◽  
Membrane Integrity ◽  
Bubalus Bubalis ◽  
Semen Parameters ◽  
Suction Pump ◽  
Bull Semen ◽  
Buffalo Semen ◽  
Live Sperm ◽  
And Control

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.

Sign in / Sign up

Export Citation Format

Share Document