Partial deoxygenation of extender improves sperm quality, reduces lipid peroxidation and reactive oxygen species during cryopreservation of buffalo (Bubalus bubalis) semen

Balamurugan, B.; Ghosh, SK; Lone, Shabir Ahmad; Prasad, J.K.; Das, G.; Katiyar, Rahul; Mustapha, Adam; Kumar, Ajay; Verma, Subhash

doi:10.1016/j.anireprosci.2017.12.008

Cited by 35 publications

(30 citation statements)

References 53 publications

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“…The better sperm freezability in terms of PM, VIB, AI, HOS responsive sperm in groups III (post‐thaw dissolved oxygen (DO): 2.38 ± 0.42 ppm) and IV (post‐thaw DO: 5.75 ± 0.48 ppm) as compared to control (post‐thaw DO: 7.98 ± 0.69) and group II (0.96 ± 0.31) was observed. Our results are in agreement with the reports of Balamurugan et al () who reported flushing extender with liquid nitrogen reduced DO levels and improved prefreeze and post‐thaw sperm quality. Previous reports have shown that degasifying extender with liquid nitrogen enhances sperm quality (Pande et al, ).…”

Section: Discussionsupporting

confidence: 93%

“…The reasons might be different assay method used for ROS, species and breed variation. Our reports are also in line with the findings of Balamurugan et al () who revealed that flushing extender with the liquid nitrogen reduced ROS and lipid peroxidation during cryopreservation of buffalo sperm.…”

Section: Discussionsupporting

confidence: 93%

“…Our reports are also in line with the findings of Balamurugan et al (2018) who revealed that flushing extender with the liquid nitrogen TA B L E 2 Effect of various levels of dissolved oxygen in extender on ROS, cholesterol (C) and phospholipid (P) at prefreeze and post-thaw stage (mean ± SEM, n = 24) reduced ROS and lipid peroxidation during cryopreservation of buffalo sperm.…”

Section: Discussionsupporting

confidence: 90%

“…Live sperm count was carried out by staining the sperm with eosin‐nigrosin. Stain preparation and method of staining have been described elsewhere (Balamurugan et al, ).…”

Section: Methodsmentioning

confidence: 99%

“…Spermatic ROS estimation was carried out using a high‐throughput spectrophotometric assay as described by Hayashi et al () with some modifications. The detailed procedure has been described elsewhere (Balamurugan et al, ).…”

Section: Methodsmentioning

confidence: 99%

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Effect of various levels of dissolved oxygen on reactive oxygen species and cryocapacitation‐like changes in bull sperm

Amin

Prasad

Ghosh

et al. 2018

Reprod Domestic Animals

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The present investigation was carried out to study the effect of various levels of dissolved oxygen (DO) on reactive oxygen species (ROS) and cryocapacitation-like changes in bull sperm. Egg yolk-Tris-glycerol (EYTG) extender was split into four subextenders; viz., Extender I (control; no flushing with liquid nitrogen (LN )), Extender II, Extender III and Extender IV were flushed with LN for 40, 16 and 8 min, respectively. The DO levels were standardized to 11.7, 2, 4 and 8 ppm, respectively, in control (Extender I), Extender II, Extender III and Extender IV. Ejaculates with mass motility of ≥ 3+ were divided into group I (diluted with Extender I), group II (diluted with Extender II), group III (diluted with Extender III) and group IV (diluted with Extender IV) up to 80 × 10 sperm/ml. Extended semen samples were packed in French mini straws (0.25 ml), equilibrated and cryopreserved. Semen samples were evaluated at prefreeze and post-thaw stage for various parameters (DO, progressive motility (PM), viability (VIB), acrosomal integrity (AI), hypo-osmotic swelling (HOS) test, ROS, cholesterol (C) and phospholipid (P). The percentage of PM, VIB, AI, HOS test, cholesterol (C) and phospholipid (P) levels, and capacitated sperm were significantly (p < 0.05) higher in groups III and IV as compared to groups I and II. However, the acrosome-reacted sperm (%; pattern AR) were significantly (p < 0.05) decreased in group III as compared to all other groups. Besides the proportion of sperm displaying tyrosine-phosphorylated pattern, EA (fluorescence at both equatorial and anterior acrosomal regions, i.e. high capacitation level) was significantly (p < 0.05) reduced in group III compared to all other groups. In conclusion, varying DO levels in the extender significantly affect sperm quality, ROS production and capacitation-like changes in bulls.

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Section: Discussionsupporting

confidence: 93%

Section: Discussionsupporting

confidence: 93%

Section: Discussionsupporting

confidence: 90%

“…Live sperm count was carried out by staining the sperm with eosin‐nigrosin. Stain preparation and method of staining have been described elsewhere (Balamurugan et al, ).…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Effect of various levels of dissolved oxygen on reactive oxygen species and cryocapacitation‐like changes in bull sperm

Amin

Prasad

Ghosh

et al. 2018

Reprod Domestic Animals

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Escherichia coli membrane‐derived oxygen‐reducing enzyme system (Oxyrase) protects bubaline spermatozoa during cryopreservation

Dalal

Chandolia

Jan

et al. 2020

Molecular Reproduction Devel

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The objective of this study was to determine the effectiveness of deoxygenation of semen extender using Escherichia coli membrane-derived oxygen scavenger (Oxyrase) on post-thaw quality of buffalo (Bubalus bubalis) spermatozoa. Sixteen semen ejaculates, four each from four bulls, were each divided into five equal fractions, diluted using Tris-egg yolk extender supplemented with different concentrations of Oxyrase (0, 0.3, 0.6, 0.9, and 1.2 U/ml), designated as treatments T1, T2, T3, T4, and T5, respectively, and cryopreserved. Immediately after thawing, Oxyrase did not improve sperm kinetics and motility; however, it improved the keeping quality (significantly lower deterioration of post-thaw sperm motility after incubation for 120 min) in T3. Further, T3 reduced (p < .05) cholesterol efflux and protected the intactness of the sperm plasma membrane. Flow cytometry with Fluo-3 AM/propidium iodide (PI) dual staining revealed the highest (p < .05) proportion of live spermatozoa with low intracellular calcium in T3. Oxyrase supplementation protected spermatozoa from premature capacitation which was confirmed by low expression of tyrosine-phosphorylated proteins (32, 75, and 80 kDa) and a relatively lower percentage of F-pattern (uncapacitated spermatozoa) in chlortetracycline assay. Importantly, the Oxyrase fortification decreased superoxide anion in a dose-dependent manner indicating reduced availability of oxygen at sperm mitochondrial level. Similarly, in Oxyrase-fortified sperm, malondialdehyde concentration, an index of lipid peroxidation, is also reduced in a dose-dependent manner. In conclusion, we demonstrate that deoxygenation of buffalo semen by Oxyrase has the potential of improving post-thaw sperm quality by overcoming the problem of cryocapacitation and oxidative damage during cryopreservation process.

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Buffalo Semen Cryopreservation: An Update

Kumar

Adnot

2022

Current Concepts in Bovine Reproduction

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Partial deoxygenation of extender improves sperm quality, reduces lipid peroxidation and reactive oxygen species during cryopreservation of buffalo (Bubalus bubalis) semen

Cited by 35 publications

References 53 publications

Effect of various levels of dissolved oxygen on reactive oxygen species and cryocapacitation‐like changes in bull sperm

Effect of various levels of dissolved oxygen on reactive oxygen species and cryocapacitation‐like changes in bull sperm

Escherichia coli membrane‐derived oxygen‐reducing enzyme system (Oxyrase) protects bubaline spermatozoa during cryopreservation

Buffalo Semen Cryopreservation: An Update

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