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  1. Fulltext Ultrastructural assessment of fresh, capacitated and acrosome-reacted Jermasia goat sperm by electron microscopy
    Hashida, N.H., Abdullah, R.B.
    ASM Science Journal, 2008;2(1):65-73.
    MyJurnal
    This study was carried out to compare the ultrastructure of fresh, capacitated and acrosome-reacted sperm. The sperm was treated with heparin for capacitation and calcium ionophore for acrosome reaction induction. Sperm samples were then prepared for ultrastructural studies and examined by transmission electron microscopy (TEM). Ultrastructural changes in plasma and acrosomal membranes, shape of the mitochondria and outer dense fibres, in capacitated and acrosome-reacted sperm were evident. The plasma membrane of fresh sperm was loosely fitted around the sperm head and the acrosomal membrane was closely opposed to the nucleus. The plasma and acrosomal membranes of the capacitated sperm were expanded, but disintegrated in the acrosome-reacted sperm. Mitochondria of fresh sperm appeared to be rounded in shape with plasma membrane closely opposed to it and the nine outer dense fibres were almost regular rounded in shape. However, in both capacitated and acrosome-reacted sperm, the mitochondria were almost regular and elongated in shape whilst the outer dense fibres were irregular in shape in the capacitated and acrosome-reacted sperm. There were no noticeable morphological changes found in the axonemal complexes in fresh, capacitated and acrosome-reacted sperm. Ultrastructural studies are able to provide detailed information on sequential events involving numerous physiological changes during fertilization.
    Matched MeSH terms: Acrosome; Acrosome Reaction
  2. Endometriosis and infertility: raised iron concentration in the peritoneal fluid and its effect on the acrosome reaction
    Arumugam K
    Hum Reprod, 1994 Jun;9(6):1153-7.
    PMID: 7962392
    Endometriosis and infertility are commonly associated. This study investigated the role of accelerated lipid peroxidation of spermatozoa by the peritoneal fluid of patients with endometriosis as a cause for this association. It proposes that the increased iron concentration present in the fluid of these patients acts as a catalyst for the process. Peritoneal fluid from 25 patients with endometriosis and 25 matched controls was obtained at laparoscopy. Spermatozoa were incubated in the fluid from both groups and the subsequent acrosome reaction rates analysed. The relationship between these results and iron concentration in the fluid was examined. A significant decrease in the acrosome reaction rate was seen in the endometriotic group (P = 0.034). Overall, a decrease in the acrosome reaction rate was associated with an increased iron concentration in the fluid (18 of the 25 pairs). In mild disease, (six of 11 pairs), the relationship was not as marked as that in severe disease (12 of 14 pairs). These results suggest that the peritoneal fluid in patients with endometriosis has a detrimental action on the acrosome reaction of spermatozoa in vitro.
    Matched MeSH terms: Acrosome/drug effects; Acrosome/physiology*
  3. Effect of butylated hydroxytoluene on cryopreservation of Boer goat semen in Tris egg yolk extender
    Memon AA, Wahid H, Rosnina Y, Goh YM, Ebrahimi M, Nadia FM, et al.
    Anim. Reprod. Sci., 2011 Nov;129(1-2):44-9.
    PMID: 22024366 DOI: 10.1016/j.anireprosci.2011.10.004
    The aim of this study was to determine the effect of butylated hydroxytoluene (BHT), a lipid-soluble anti-oxidant added in different concentrations to the Tris egg yolk extenders on semen cytological parameters pre freezing and post thawing (motility, morphology, viability, acrosome integrity and membrane integrity) of Boer goat spermatozoa. A total of 40 ejaculates from four Boer goat bucks were collected using an artificial vagina. Ten replicates of the ejaculates were diluted with a Tris egg yolk based extender which contained various concentrations (0.5mM, 1.0mM, 2.0mM and 3.0mM) of butylated hydroxytoluene while one sample was processed without supplementation of antioxidant and served as control. The diluted semen was cooled at 4°C and loaded into the straw and then stored in liquid nitrogen. It was evident that supplementation of BHT produces positive effect in terms of motility, membrane integrity and acrosome integrity in comparison with the control group in cooled and frozen Boer goat semen. Results showed significant differences in motility, membrane integrity, acrosome integrity and viability of cooled and frozen Boer goat spermatozoa at different concentrations. Motility, membrane integrity, acrosome integrity and viability was significantly higher in all treated groups than the control group (P<0.05) while there was no significant differences (P>0.05) in morphology trait between all group in cooled semen. However, improvement (P<0.05) was observed only in terms of the membrane integrity and acrosome integrity compared to the control and other treated groups in frozen semen. In conclusion, BHT can be used in cryopreservation of Boer goat semen in order to reduce the oxidative stress on spermatozoa.
    Matched MeSH terms: Acrosome/physiology; Acrosome/ultrastructure
  4. Effect of docosahexanoic acid on quality of frozen-thawed bull semen in BioXcell extender
    Kaka A, Haron W, Yusoff R, Yimer N, Khumran AM, Sarsaifi K, et al.
    Reprod Fertil Dev, 2017 Mar;29(3):490-495.
    PMID: 28442061 DOI: 10.1071/RD15089
    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 15ngmL(-1) DHA was added. The supplemented semen samples were incubated at 37°C for 15min for DHA uptake by spermatozoa. Later, samples were cooled for 2h at 5°C and packaged into 0.25-mL straws, frozen in liquid nitrogen for 24h and subsequently thawed for evaluation. Results are presented as percentages ± s.e.m. Supplementation with DHA at 3ngmL(-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, 3ngmL(-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.
    Matched MeSH terms: Acrosome
  5. Fulltext Effect of different concentrations of soybean lecithin and virgin coconut oil in Tris-based extender on the quality of chilled and frozen-thawed bull semen
    Tarig AA, Wahid H, Rosnina Y, Yimer N, Goh YM, Baiee FH, et al.
    Vet World, 2017 Jun;10(6):672-678.
    PMID: 28717321 DOI: 10.14202/vetworld.2017.672-678
    AIM: The objective of this study was to evaluate the effects of different concentrations of soybean lecithin (SL) and virgin coconut oil (VCO) in Tris-based extender on chilled and frozen-thawed bull semen quality parameters.

    MATERIALS AND METHODS: A total of 24 ejaculates were collected from four bulls via an electroejaculator. Semen samples were diluted with 2% VCO in Tris-based extender which consists of various concentrations of SL (1, 1.25, 1.5, and 1.75%). A 20% egg yolk in Tris used as a positive control (C+). The diluted semen samples were divided into two fractions; one for chilling which were stored at 4°C for 24, 72, and 144 h before evaluated for semen quality parameters. The second fraction used for freezing was chilled for 3 h at 4°C, packed into 0.25 mL straws and then cryopreserved in liquid nitrogen. The samples were then evaluated after 7 and 14 days. Chilled and frozen semen samples were thawed at 37°C and assessed for general motility using computer-assisted semen analysis, viability, acrosome integrity and morphology (eosin-nigrosin stain), membrane integrity, and lipid peroxidation using thiobarbituric acid reaction test.

    RESULTS: The results showed that all the quality parameters assessed were significantly (p<0.05) improved at 1.5% SL concentration in chilled semen. Treatment groups of 1, 1.25, 1.5, and 1.75% SL were higher in quality parameters than the control group (C+) in chilled semen. However, all the quality parameters in frozen-thawed semen were significantly higher in the C+ than the treated groups.

    CONCLUSION: In conclusion, supplementation of 1.5% SL in 2% VCO Tris-based extender enhanced the chilled bull semen. However, there was no marked improvement in the frozen-thawed quality parameters after treatment.

    Matched MeSH terms: Acrosome
  6. Two-dimensional polyacrylamide gel electrophoresis of Bali bull (Bos javanicus) seminal plasma proteins and their relationship with semen quality
    Sarsaifi K, Haron AW, Vejayan J, Yusoff R, Hani H, Omar MA, et al.
    Theriogenology, 2015 Oct 1;84(6):956-68.
    PMID: 26119476 DOI: 10.1016/j.theriogenology.2015.05.035
    The present study evaluated the relationship between Bali bull (Bos javanicus) seminal plasma proteins and different semen quality parameters. Semen samples from 10 mature Bali bulls were evaluated for conventional semen parameters (general motility, viability, and normal morphology), sperm functionality (acrosome reaction, sperm penetration rate, sperm penetration index), sperm kinetics (computer-assisted semen analysis parameters such as sperm velocity), and sperm morphology (acrosome and membrane integrity). Frozen-thawed semen with higher sperm motility, viability, acrosome integrity, and membrane integrity (P < 0.05) are consistently higher in acrosome reaction and sperm penetration assay. Three bulls showed the highest, four bulls displayed the medium, and the remaining three bulls showed the lowest for all sperm parameters and SPA. The proteome maps of seminal plasma from high-quality and low-quality Bali bulls were also established. Seminal plasma of both high-quality and low-quality Bali bulls was subjected to two-dimensional SDS-PAGE with isoelectric point ranged from 3 to 10 and molecular weight from 10 to 250 kDa. Approximately 116 spots were detected with Blue Silver stain, and of these spots, 29 were selected and identified by MALDI-TOF/TOF-MS/MS. A majority of the proteins visualized in the seminal plasma two-dimensional maps was successfully identified. An essential group of the identified spots represented binder of sperm 1 (BSP1), clusterin, spermadhesin, tissue inhibitor of metalloproteinases 2 (TIMP-2), and phospholipase A2 (PLA2). Other proteins found in high abundance included seminal ribonuclease, serum albumin, cationic trypsin, and peptide similar to β2 microglobulin. Thus, a reference map of Bali bull seminal plasma proteins has been generated for the very first time and can be used to relate protein pattern changes to physiopathologic events that may influence Bali bull reproductive performance.
    Matched MeSH terms: Acrosome; Acrosome Reaction
  7. Fulltext α-Linolenic acid supplementation in BioXcell® extender can improve the quality of post-cooling and frozen-thawed bovine sperm
    Kaka A, Wahid H, Rosnina Y, Yimer N, Khumran AM, Sarsaifi K, et al.
    Anim. Reprod. Sci., 2015 Feb;153:1-7.
    PMID: 25544152 DOI: 10.1016/j.anireprosci.2014.12.001
    The present study was conducted to determine the effects of supplementing α-linolenic acid (ALA) into BioXcell(®) extender on post-cooling, post-thawed bovine spermatozoa and post thawed fatty acid composition. Twenty-four semen samples were collected from three bulls using an electro-ejaculator. Fresh semen samples were evaluated for general motility using computer assisted semen analyzer (CASA) whereas morphology and viability with eosin-nigrosin stain. Semen samples extended into BioXcell(®) were divided into five groups to which 0, 3, 5, 10 and 15 ng/ml of ALA were added, respectively. The treated samples were incubated at 37°C for 15 min for ALA uptake by sperm cells before being cooled for 2 h at 5°C. After evaluation, the cooled samples were packed into 0.25 ml straws and frozen in liquid nitrogen for 24 h before thawing and evaluation for semen quality. Evaluation of cooled and frozen-thawed semen showed that the percentages of all the sperm parameters improved with 5 ng/ml ALA supplement. ALA was higher in all treated groups than control groups than control group. In conclusion, 5 ng/ml ALA supplemented into BioXcell(®) extender improved the cooled and frozen-thawed quality of bull spermatozoa.
    Matched MeSH terms: Acrosome/drug effects
  8. Effect of sugars on characteristics of Boer goat semen after cryopreservation
    Naing SW, Wahid H, Mohd Azam K, Rosnina Y, Zuki AB, Kazhal S, et al.
    Anim. Reprod. Sci., 2010 Oct;122(1-2):23-8.
    PMID: 20637550 DOI: 10.1016/j.anireprosci.2010.06.006
    In order to improve Boer goat semen quality during cryopreservation process, the influence of sugar supplementation on semen characteristics of sperm were investigated. Three experiments were carried out to investigate the effect of (a) addition of two monosaccharides (fructose and glucose) and two disaccharides sugars (trehalose and sucrose) (b) sugar combination (fructose and trehalose, sucrose and trehalose, glucose and trehalose), and control (glucose without trehalose) (c) different concentrations of trehalose on cryopreservation using Tris based extender. The total motility, forward motility, viability, normal spermatozoa, acrosome integrity and membrane integrity were assessed subjectively. Differences were not detected among monosaccharides, but glucose increased (P<0.05) sperm forward motility in post-thaw goat semen compared to trehalose or sucrose supplementation. Semen quality did not differ (P>0.05) among disaccharide sugar supplementation. Combination of glucose and trehalose significantly improved the characteristics of Boer spermatozoa after cryopreservation (P<0.05). Supplementation of trehalose (198.24mM) into the glucose extender significantly increased total motility, forward motility, live spermatozoa, acrosome integrity and membrane integrity following cryopreservation (P<0.05). In conclusion, glucose had the better ability to support Boer sperm motility and movement patterns. Combination of monosaccharide (glucose) and disaccharide (trehalose) improved semen quality following cryopreservation. Trehalose supplementation at the concentration of 198.24mM to the glucose extender conferred the greater improvement of semen quality for Boer semen cryopreservation.
    Matched MeSH terms: Acrosome/drug effects
  9. Fulltext First Evaluations and Cryopreservation of Semen Samples from Sunda Clouded Leopards (Neofelis diardi)
    Zainuddin ZZ, Mohamed Tarmizi MR, Yap KC, Comizzoli P, Sipangkui S
    Animals (Basel), 2020 06 22;10(6).
    PMID: 32580372 DOI: 10.3390/ani10061072
    A better understanding of semen characteristics and resilience to freezing temperatures is necessary before developing assisted reproductive techniques and systematic biobanking for the Sunda clouded leopard. The objective of this study was to evaluate for the first time the semen and sperm quality (in fresh and frozen samples) of two captive Sunda clouded leopards in Malaysia. A total of 17 examinations of the reproductive tract (using ultrasonography) and electro-ejaculations were performed on the two leopards over a 2-year period. Samples obtained from Leopard 1 (8 years old) varied in terms of volume (402 ± 92 µL), pH (7.9 ± 0.9), sperm motility (54.5 ± 24.2%), sperm concentration (122.4 ± 84.7 × 106 sperm/mL), normal morphology (23.9 ± 12.3%), and viability (55.2 ± 18.2%). Midpiece defects represented the most common structural abnormality followed by abnormal tail and head defects. Samples from Leopard 2 (11 year old with abnormal testicular tissue) were of lesser quality. Two frozen semen samples from Leopard 1 were thawed and examined for acrosome integrity. Post-thawed samples contained <10% of motile spermatozoa but almost 50% of abnormal acrosomes. The present results emphasized the high incidence of structurally-abnormal spermatozoa, similar to the mainland clouded leopard. Post-thaw evaluations showed that the few surviving spermatozoa could potentially be used for in vitro fertilization or sperm injection. However, more individuals must be studied to validate those first findings that are exciting but still preliminary.
    Matched MeSH terms: Acrosome
  10. Oxidative stress and sperm function: A systematic review on evaluation and management
    Dutta S, Majzoub A, Agarwal A
    Arab J Urol, 2019;17(2):87-97.
    PMID: 31285919 DOI: 10.1080/2090598X.2019.1599624
    Objective: To review and present the most distinct concepts on the association of reactive oxygen species (ROS) with male reproduction. Methods: The Preferred Reporting Items for Systematic Reviews and Meta Analyses (PRISMA) guidelines were used to search PubMed, Medline, EMBASE, and the Cochrane electronic databases for studies investigating the role of oxidative stress (OS) on sperm function. Results: The literature search yielded 1857 studies, of which 1791 articles were excluded because of irrelevance of data, non-English language, non-human nature or because they were case reports or commentaries. All included studies were reviews (46), meta-analyses (one), original research studies (18) and guideline articles (one). The studies were published between 1984 and 2018. Under normal physiological conditions, ROS are vital for sperm maturation, hyperactivation, capacitation, acrosome reaction, as well as fertilisation. However, a number of endogenous and exogenous causes may induce supra-physiological levels of ROS resulting in lipid peroxidation, sperm DNA fragmentation and apoptosis, and consequently infertility. Several laboratory testing methods can be used in infertile men to diagnose OS. Treatment usually involves antioxidant supplementation and, when possible, elimination of the causative factor. Conclusion: OS is an important cause of male factor infertility. Its assessment provides essential information that can guide treatment strategies aimed at improving the male's reproductive potential. Abbreviations: bp: base-pair; CAT: catalase; LPO: lipid peroxidation; MDA: malondialdehyde; MiOXSYS: Male Infertility Oxidative System; mtDNA: mitochondrial DNA; NAD(PH): nicotinamide adenine dinucleotide (phosphate); NO: nitric oxide; 8-OHdG: 8-hydroxy-2'-deoxyguanosine; ORP: oxidation-reduction potential; OS: oxidative stress; PKA: protein kinase A; PLA2: phospholipase A2; PRISMA: Preferred Reporting Items for Systematic Reviews and Meta-Analyses; PUFA: poly-unsaturated fatty acid; ROS: reactive oxygen species; SOD: superoxide dismutase; TAC: total antioxidant capacity; TBA: thiobarbituric acid.
    Matched MeSH terms: Acrosome Reaction
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