Reduction of dissolved oxygen in semen extender with nitrogen gassing reduces oxidative stress and improves post-thaw semen quality of bulls

Sperm cryopreservation is an assisted reproductive technique routinely used in canine species for genetic conservation. However, during cryopreservation, the DNA damages are still elevated, limiting the fertilization rate. The present study was conducted to evaluate whether supplementation of canine semen extender with a molecule limiting the metabolic activities can improve the quality of frozen-thawed canine spermatozoa. We used metformin, known to limit the mitochondrial respiratory and limit the oxidative stress. Before and during the freezing procedure, metformin (50µM and 500µM) has been added to the extender. After thawing, sperm exposed to metformin conserved the same viability without alteration in the membrane integrity or acrosome reaction. Interestingly, 50µM metformin improved the sperm motility in comparison to the control, subsequently increasing mitochondrial activity and NAD+ content. In addition, the oxidative stress level was reduced in sperm treated with metformin improving the sperm quality as measured by a different molecular marker. In conclusion, we have shown that metformin is able to improve the quality of frozen-thawed dog semen when it is used during the cryopreservative procedure.

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Identifikasi Bakteri dan Efektivitas Antibiotik dalam Pengencer untuk Mengontrol Pertumbuhan Bakteri pada Semen Sapi Friesian Holstein

jurnal veteriner ◽

10.19087/jveteriner.2019.20.1.140 ◽

2019 ◽

Vol 20 (1) ◽

pp. 140

Author(s):

Muttaqinullah Rabusin ◽

Andriani Andriani ◽

Raden Iis Arifiantini ◽

Ni Wayan Kurniani Karja

Keyword(s):

Semen Quality ◽

Bacterial Species ◽

Enterobacter Cloacae ◽

Bovine Semen ◽

Frozen Semen ◽

Bacterial Content ◽

Pseudomonas Diminuta ◽

Semen Extender ◽

Fresh Semen

This study was designed to investigate the presence of bacterial species in Friesian Holstein (FH) bovine semen at the time of collection, processing and to assess the efficacy of two types of antibiotics combinations; penicillin and streptomycin (PS) and gentamycin, tylosin, lincomycin and spectinomycin (GTLS) in semen extender on bacterial control and quality of semen. For this purpose, three experiments were conducted. In experiment 1, identification of bacterial content in fresh semen which collected from 5 bovine ejaculates. In experiment 2, identification of bacterial content in skimmilk-eggyolk extender which were prepared in artificial insemination center, Lembang, Bandung. In experiment 3, identification of bacterial content in frozen thawed semen. In the result, some of bacterial species were isolated from the bovine semen. The GTLS combination of antibiotics may be incorporated into a freezing extender or protocol without compromising the post-thawed semen quality of FH bull spermatozoa. Three types of bacteria were found in fresh semen; Klebsiella sp., Micrococcus sp., and Pediococcus sp.. Three types of bacteria were found in semen extender; Enterobacter cloacae, Pseudomonas diminuta and Serratia plymutica. Two types of bacteria were found in frozen semen; Enterobacter cloacae and Serratia plymutica. In conclusion, antibiotics PS and GTLS were effective for controlling the growth of bacteria in frozen semen.

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PSV-9 Effects of Natural Honey Inclusion in Dilution and Freezing Extenders on Frozen-thawed Semen Quality in Boars

Journal of Animal Science ◽

10.1093/jas/skab054.346 ◽

2021 ◽

Vol 99 (Supplement_1) ◽

pp. 211-211

Author(s):

Kayode B Balogun ◽

Griffin Nicholls ◽

Olujide Sokunbi ◽

Kara R Stewart

Keyword(s):

Semen Quality ◽

Swine Industry ◽

Normal Cells ◽

Progressive Motility ◽

Normal Sperm ◽

Frozen Semen ◽

Boar Semen ◽

Semen Extender ◽

Natural Honey

Abstract Improvements in the post-thaw quality of frozen semen could allow increased utilization of this technology in the swine industry. The objective of this study was to investigate the effects of natural honey inclusion in semen extender and freezing media on motility, mobility, and morphology of cryopreserved boar sperm. Ejaculates from 6 terminal cross-bred boars were collected using the gloved-hand technique for 3 weeks and used in a 2 x 3 factorial study design. Following collection, semen samples were incubated overnight in dilution extender with and without natural honey (D0: Androhep Plus; D1: Androhep Plus + 0.25% honey). The following day, the semen samples were cooled to 4 C in LEY cooling extender then frozen in freezing media containing 93% cooling extender + 6% glycerol + 1% Equex-STM Paste (F1), or freezing media with natural honey replacing 50% of the glycerol (F2) or 50% of the Equex-STM paste (F3). Semen samples were frozen using a controlled-rate freezer and stored in liquid nitrogen. Two straws per treatment for each boar were thawed and semen quality assessed. The inclusion of natural honey in dilution extender had no effect on post-thaw motility (P=0.733), progressive motility (P=0.562), or other mobility parameters (0.995≤P≥0.081). However, D1 had a higher percentage of normal acrosomes (P=0.001) and morphologically normal cells (P< 0.001) resulting from lower tail abnormalities compared to D0 (P=0.006). Post-thaw motility (P< 0.001) and progressive motility (P< 0.001) were increased in F3 compared to both F2 and F1. F1 had reduced normal acrosomes (P=0.009) and morphologically normal cells (P< 0.001) resulting from higher tail abnormalities (P< 0.001). In conclusion, the inclusion of natural honey, at 0.25%, in dilution extender helps maintain normal sperm and acrosome morphology, and replacing 50% Equex-STM Paste with honey in freezing extender improves post-thaw sperm motility and progressive motility of frozen-thawed boar semen.

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Role of α-Tocopherol and β-Carotene in Ameliorating the Fenvalerate-Induced Changes in Oxidative Stress, Hemato-Biochemical Parameters, and Semen Quality of Male Rats

Journal of Environmental Science and Health Part B ◽

10.1081/pfc-120035929 ◽

2004 ◽

Vol 39 (3) ◽

pp. 443-459 ◽

Cited By ~ 74

Author(s):

F. M. El-Demerdash ◽

M. I. Yousef ◽

F. S. Kedwany ◽

H. H. Baghdadi

Keyword(s):

Oxidative Stress ◽

Biochemical Parameters ◽

Semen Quality ◽

Male Rats ◽

Induced Changes ◽

Β Carotene

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Maternal dietary supplementation with grape seed extract in reproductive hens increases fertility in females but decreases semen quality in males of the F1 generation

PLoS ONE ◽

10.1371/journal.pone.0246750 ◽

2021 ◽

Vol 16 (2) ◽

pp. e0246750

Author(s):

Jérémy Grandhaye ◽

François Lecompte ◽

Pascal Chartrin ◽

Maryse Leconte ◽

Antonella Riva ◽

...

Keyword(s):

Oxidative Stress ◽

Egg Quality ◽

Semen Quality ◽

Grape Seed Extract ◽

Grape Seed ◽

Seed Extract ◽

The Body ◽

Male And Female ◽

F1 Generation

Genetic selection in parental broiler breeders has increased their susceptibility to metabolic disorders and reproductive dysfunction. We have recently shown that maternal dietary grape seed extract (GSE) supplementation in hens improves fertility parameters, egg quality, oxidative stress in different tissues and the quality of F1 chicks. Here, we analysed the growth and fertility (both female and male) of the F1 generation animals and the quality of their offspring (F2 generation). Eggs issued from hens supplemented with GSE presented lower ROS production than control hens, suggesting a change in the embryonic environment. However, this did not affect the growth nor the body composition of male and female F1s from hatching to adulthood (37 weeks of age). At 37 weeks of age, the biochemistry analysis of the GSE-F1 muscle has revealed an increase in sensitivity to oxidative stress and a slight change in lipid composition. Both male and female F1-GSE groups presented a delay in puberty with a lower testis volume at 30 weeks of age and lower ovary development at 26 weeks of age. Adult GSE-F1 males did not present histological alterations of seminiferous tubules or semen production, but the semen quality was degraded due to higher oxidative stress and DNA-damaged spermatozoa compared with control F1 animals. In adult GSE-F1 females, despite the delay in puberty, the females laid more eggs of better quality (fewer broken eggs and a higher hatching rate). At hatching, the weight of the chicks from GSE-F1 females was reduced, and this effect was stronger in F2 male chicks (F2) compared with F2 control chicks (F2), because of the lower muscle volume. In conclusion, we can raise the hypothesis that maternal dietary GSE supplementation produces eggs with change in embryonic metabolism, which may affect in adulthood the fertility. The data obtained from the F1-GSE group pointed to a sex-specific modification with higher egg quality in females but semen sensitive to stress in males. Finally, male F2 chicks were leaner than control chicks. Thus, maternal dietary grape seed extract (GSE) supplementation in hens may impact on the fertility of the offspring in a sex-specific manner in subsequent generations.

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Optimal inclusion level of butylated hydroxytoluene in semen extender improves the quality of post-thawed canine sperm

Czech Journal of Animal Science ◽

10.17221/6273-cjas ◽

2012 ◽

Vol 57 (No. 8) ◽

pp. 377-381 ◽

Cited By ~ 5

Author(s):

M. Ziaullah ◽

A. Ijaz ◽

M. Aleem ◽

A.K. Mahmood ◽

H. Rahman ◽

...

Keyword(s):

Semen Quality ◽

Membrane Integrity ◽

Egg Yolk ◽

Quality Parameters ◽

Butylated Hydroxytoluene ◽

Osmotic Swelling ◽

Inclusion Level ◽

Semen Extender ◽

Supravital Staining

The study was conducted to evaluate the potential cryoprotective effect of butylated hydroxytoluene (BHT) through post-thaw evaluation of canine semen and its optimal inclusion level. Ejaculated canine semen was extended in TRIS-glucose egg yolk extender containing various concentrations of BHT (0.5, 1.0, 1.5, 2.0, and 2.5mM). Semen was frozen at −196°C using 200 × 10<sup>6 </sup>spermatozoa per 0.5 ml straws and post-thaw evaluation was carried out in terms of sperm motility, viability, plasma membrane integrity, and acrosomal integrity through phase-contrast microscope, supravital staining, hypo-osmotic swelling test, and normal acrosomal ridge, respectively. BHT was found to improve (P > 0.005) all post-thawed semen quality parameters at an inclusion level of 1.0mM in the extended semen. However, higher concentrations than this were found to have detrimental effects.  

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Effect of optimization of the levels of dissolved oxygen in semen extender on physico-morphological attributes and functional membrane integrity of crossbred bull spermatozoa

Indian Journal of Animal Research ◽

10.18805/ijar.b-3585 ◽

2018 ◽

Author(s):

Abhishek Kumar ◽

J. K. Prasad ◽

A. R. Mustapha ◽

R. Mustapha ◽

G. K. Das ◽

...

Keyword(s):

Dissolved Oxygen ◽

Semen Quality ◽

Membrane Integrity ◽

Control Group ◽

Morphological Attributes ◽

Progressive Motility ◽

Group I ◽

Functional Membrane ◽

Group Ii ◽

Semen Extender

The objective of present study was to investigate the effects of two different levels of dissolved oxygen (DO) i.e. 4 and 8 ppm in the extenders on physico-morphological attributes and functional membrane integrity of crossbred bull spermatozoa following cryopreservation. Only those ejaculates that had a mass activity of ³ 3+ and individual progressive motility of ³ 70% were utilized. Each of the ejaculates were divided and extended to have 80×106 spermatozoa/ mL of extender in three groups i.e. Group I (control), Group II (4 ppm) and Group III (8 ppm). Progressive motility (%) was higher (p Lass Than 0.05) in Group II and III as compared to Group I at both pre-freeze and post-thaw stage. Per cent live sperm and HOST reactive sperm were higher (p Lass Than 0.05) in Group II and in Group II and III as compared to Group I at pre-freeze and post-thaw stage, respectively. Sperm abnormality (%) was lower (p Lass Than 0.05) in Group II at post-thaw stage. Per cent acrosome intact sperm did not differ (p>0.05) among the groups at pre-freeze stage, but, were higher (p Lass Than 0.05) in Group II and III as compared to Group I at post-thaw stage. In conclusion, cryopreservation of semen having DO level of 4 ppm in semen extender yielded better results followed by 8 ppm in terms of semen quality parameters and functional membrane integrity of crossbred bull spermatozoa.

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