Manganese Provides Antioxidant Protection for Sperm Cryopreservation that May Offer New Consideration for Clinical Fertility

Reactive oxygen species (ROS) are generated by sperm metabolism. While, ROS are required for maturation, capacitation and acrosome reaction, they also modify many peroxidable cellular compounds. There is production of ROS during cryopreservation and frozen spermatozoa are highly sensitive to lipid peroxidation (LPO). Antioxidants exert a protective effect on the plasma membrane of frozen bovine sperm preserving both metabolic activity and cellular viability. Manganese (Mn++) is proved to be a chain breaking antioxidant in biological system. Therefore, we examined the role of (Mn++) during cryopreservation of cattle bull semen. Semen was divided into four parts and cryopreserved in egg-yolk-citrate extender + glycerol (EYC-G), EYC-G + 100 µM of Mn++, EYC-G + 150 µM of Mn++and EYC-G + 200 µM of Mn++. After four hours of cooling and 24 hrs of freezing, the spermatozoa were examined for percentage motility, Hypo-osmotic swelling (HOS), LPO and protein leakage. Addition of manganese to the semen during cryopreservation showed a protective effect and accounted for an increase in semen quality parameters [percentage motility, HOS percent and decrease in malondialdehyde (MDA) production and protein leakage]. The effect of manganese on motility and HOS was non-significant (p < 0.05) in cooled spermatozoa but significant with 150 µM of Mn++in frozen-thawed spermatozoa. MDA production and protein leakage decreased to a significant and maximum level (p < 0.05) on addition of 200 µM of manganese. The addition of manganese to EYC-G dilutor will improve the quality/fertility of semen, which will result in improvement of in vitro fertilization and artificial insemination success rate.

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Effect of whole and clarified egg yolk based extenders on post-thaw semen quality in bulls.

10.31220/agrirxiv.2021.00050 ◽

2021 ◽

Author(s):

Prosper Kamusasa ◽

Eddington Gororo ◽

Fungayi Primrose Chatiza

Keyword(s):

Semen Quality ◽

Sperm Quality ◽

Egg Yolk ◽

Quality Parameters ◽

Normal Morphology ◽

Semen Cryopreservation ◽

Bull Semen ◽

Inclusion Level ◽

Whole Egg

Abstract This study was conducted to evaluate the comparative cryoprotective effects of whole egg yolk and clarified egg yolk on post thaw sperm quality parameters and to determine the optimum clarified egg yolk inclusion level (10-20%) in semen extenders for Mashona bull semen cryopreservation. It was shown that there was a significant decrease in sperm quality variables following cryopreservation. Semen quality increased with the concentration of clarified egg yolk, indicating a positive relationship between egg yolk LDL concentration and maintenance of in vitro sperm quality. The 20% clarified egg yolk (CEY20) extender treatment gave post-thaw motility, viability and normal morphology values which were comparable to the control (20% whole egg yolk, WEY20). The 10% clarified egg yolk concentration gave the least post-thaw quality values and the greatest proportion of defective spermatozoa. This experiment found no advantage of replacing whole egg yolk with up to 15% clarified egg yolk in Mashona bull semen cryopreservation. However, 20% clarified and 20% whole egg yolk performed similarly in the maintenance of post-thaw sperm motility, viability and normal morphology.

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The Re-Addition of Seminal Plasma after Thawing Does Not Improve Buck Sperm Quality Parameters

Animals ◽

10.3390/ani11123452 ◽

2021 ◽

Vol 11 (12) ◽

pp. 3452

Author(s):

Uchechi Linda Ohaneje ◽

Uchebuchi Ike Osuagwuh ◽

Manuel Alvarez-Rodríguez ◽

Iván Yánez-Ortiz ◽

Abigail Tabarez ◽

...

Keyword(s):

Seminal Plasma ◽

Sperm Quality ◽

Membrane Integrity ◽

Egg Yolk ◽

Quality Parameters ◽

Mitochondrial Activity ◽

Dna Integrity ◽

Vitro Fertilization

In order to achieve a higher post-thaw buck sperm quality, an approach in the thawing protocol of cryopreserved sperm doses under in vitro capacitation conditions mimicking the in vivo female environment was studied. Therefore, functional and kinetic characteristics of buck thawed sperm from males of different ages, the season of collection, and melatonin implanted males in the non-breeding season were assessed after 3 h of incubation in an in vitro fertilization (IVF) media with 20% of buck seminal plasma (SP). Previously, fresh ejaculates were collected via artificial vagina from eight males of the Cabra Blanca de Rasquera breed during two consecutive years in breeding and non-breeding periods. Prior to semen collection in non-breeding seasons, males were split into two groups: one group was implanted with melatonin, while the other was not. In each group, semen samples were pooled, centrifuged, and diluted in an extender containing 15% powdered egg yolk and 5% glycerol before freezing. After thawing, sperm were washed and incubated in three different media: (a) control media (modified phosphate-buffered saline (PBS), (b) IVF commercial media, and (c) IVF media + 20% SP. Sperm motility was evaluated by CASA, while plasma and acrosome membrane integrity, mitochondria activity, and DNA fragmentation were analysed by flow cytometer at 0 h and after 3 h incubation. A significant reduction in motility, mitochondrial activity, plasma, and acrosome membrane integrity were observed after incubation in the presence of SP, although similar to that observed in IVF media alone. DNA integrity was not affected under in vitro capacitation conditions, regardless of SP addition. In conclusion, the addition of SP failed to improve post-thaw buck sperm quality under in vitro conditions irrespective of male age, the season of collection, and melatonin implant.

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Study on the in vitro fertilizing potential of mithun (Bos frontalis) semen

Indian Journal of Animal Research ◽

10.18805/ijar.11323 ◽

2016 ◽

Author(s):

P. Perumal ◽

S. K. Srivastava ◽

K. K. Baruah ◽

S. K. Ghosh

Keyword(s):

Semen Quality ◽

Egg Yolk ◽

Forward Direction ◽

Poor Quality ◽

Quality Parameters ◽

The Poor ◽

Acrosomal Membrane ◽

Sperm Abnormality ◽

Biochemical Profiles

Semen quality parameters (SQPs) such as motility, viability, total sperm abnormality, integrity of acrosome, plasma membrane and nucleus and vanguard distance travelled by sperm and biochemical profiles such as aspartate amino transaminase (AST), alanine amino transaminase (ALT) and lactate dehydrogenase (LDH) were studied in mithun at post thawed stage. Fifty ejaculates were collected through transrectal massage method from matured bulls and the ejaculates were divided into good and poor quality. The ejaculates were extended with Tris egg yolk citrate glycerol extender and stored in liquid nitrogen. PT seminal, biochemical profiles and zona binding per cent (BP) and binding index (BI) were studied with standard protocols. The results revealed good quality ejaculates has significantly (p<0.05) higher SQPs, BP & BI and has significantly (p<0.05) lower total sperm abnormality, AST, ALT and LDH than in the poor quality ejaculates. PT motility was positively (p<0.05) correlated with SQPs, vanguard distance, BI & BP and negatively correlated (p<0.05) with total sperm abnormality and biochemical profiles. In conclusion, the good quality sperm has higher functional sperm structures and higher intactness of sperm plasma and acrosomal membrane to motile faster in forward direction to reach the site of fertilization and successful fertilization.

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Protective effect of vitamin C and lycopene on the in vitro fertilization capacity of sex‐sorted bull sperm by inhibiting the oxidative stress

Reproduction in Domestic Animals ◽

10.1111/rda.13748 ◽

2020 ◽

Vol 55 (9) ◽

pp. 1103-1114

Author(s):

Pei‐Pei Zhang ◽

Jing‐Jing Wang ◽

Chong‐Yang Li ◽

Hai‐sheng Hao ◽

Hao‐Yu Wang ◽

...

Keyword(s):

Oxidative Stress ◽

In Vitro Fertilization ◽

Vitamin C ◽

Protective Effect ◽

Vitro Fertilization ◽

Bull Sperm ◽

Fertilization Capacity

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Acrosin activity in human spermatozoa in relation to semen quality and in-vitro fertilization

Human Reproduction ◽

10.1093/oxfordjournals.humrep.a138033 ◽

1993 ◽

Vol 8 (2) ◽

pp. 253-257 ◽

Cited By ~ 39

Author(s):

C.J. De Jonge ◽

S.M. Tarchala ◽

R.G. Rawlins ◽

Z. Binor ◽

E. Radwanska

Keyword(s):

In Vitro Fertilization ◽

Semen Quality ◽

Human Spermatozoa ◽

Vitro Fertilization ◽

Acrosin Activity

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Resilience, Perceived Stress, and Depressed Mood in Women Under in Vitro Fertilization Treatment

Reproductive Sciences ◽

10.1007/s43032-021-00685-1 ◽

2021 ◽

Author(s):

Carmen Fernandez-Ferrera ◽

David Llaneza-Suarez ◽

Daniel Fernandez-Garcia ◽

Vanesa Castañon ◽

Cristina Llaneza-Suarez ◽

...

Keyword(s):

Protective Effect ◽

Perceived Stress ◽

Depressed Mood ◽

Sleep Disturbances ◽

Depression Scale ◽

Short Version ◽

Cross Sectional ◽

Vitro Fertilization ◽

Infertile Women

AbstractIt has been suggested that women who display higher resilience levels may have less psychological distress during IVF. The aim of this study was to evaluate how infertile women deal with perceived stress, depressed mood, and sleep disturbances at the first IVF attempt and after one or more negative IVF outcomes depending on their level of resilience. An observational, cross-sectional study was carried out in a sample of 207 infertile women undergoing IVF procedures. The participants completed the short version of the Connor-Davidson Resilience Scale (CD-RISC), the short version of the European Spanish Version of Perceived Stress Scale (PSS-10), the Center of Epidemiologic Studies Depression Scale (CESD-10), and the Jenkins Sleep Scale (JSS). The relationship between CD-RISC scores ranked according to percentiles and mean PSS-10 scores revealed that women with strong resilience had lower perceived stress. After splitting the sample according to CD-RISC percentiles, differences were observed only at the first IVF attempt and the observed protective effect of high resilience scores appears to disappear following a negative IVF outcome. Women with high resilience are less likely to suffer from perceived stress or depressed mood during their first IVF attempt, this protective effect appears to be lost after a negative outcome.

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Effect of Age and Frequency of Collection on Quality of Jersey Bulls Semen at National Livestock Breeding Center (NLBC), Nepal

International Journal of Applied Sciences and Biotechnology ◽

10.3126/ijasbt.v7i1.23312 ◽

2019 ◽

Vol 7 (1) ◽

pp. 88-95 ◽

Cited By ~ 2

Author(s):

S. Sankhi ◽

K.R. Sapkota ◽

B. Regmi

Keyword(s):

Semen Quality ◽

Sperm Concentration ◽

Quality Parameters ◽

Bull Semen ◽

Livestock Breeding ◽

Abnormal Sperm ◽

Breeding Center ◽

Live Sperm ◽

Level Of Significance ◽

Effect Of Age

Objective: To determine the effect of age and frequency of collection on quality parameters of Jersey bull semen at National Livestock Breeding Center (NLBC), Pokhara. Materials and Methodology: Nine Jersey bulls were selected randomly from NLBC. To obtain the effect of age, six bulls of three different age interval (3-4, 5-7 and 8-9 years) were selected randomly. Remaining three bulls were used to determine the effect of frequency of collection on semen quality. Studies were conducted for three months (Nov/Dec to Dec/Jan, 2016). Semen quality was analyzed using CASA system and SMILE software. ANOVA and paired t- tests were applied to find out level of significance. All the statistical analyses were done using SPSS 17.0 Results: Sperm concentration, post- thaw motility, live sperm, and abnormal sperm were significantly affected by the age of the bulls. Semen quality was found superior in the bulls belonging to 5-7 years of age intervals. Significantly higher value for semen volume, sperm concentration and post- thaw motility was obtained on second collection (P<0.05). However, volume initial motility, pre-filling motility, live sperm and abnormal sperm percentage did not differ significantly among different age intervals and between the frequencies of collection. Conclusion: Different quality parameters of semen like volume, concentration, initial motility, pre-filling motility, post thaw motility, live and normal spermatozoa percentages were higher in 5-7 years old bulls and in second collection of semen. Int. J. Appl. Sci. Biotechnol. Vol 7(1): 88-95

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284 COMPARATIVE FERTILITY OF FRESHLY-COLLECTED VERSUS FROZEN - THAWED SPERMATOZOA FOR IN VITRO FERTILIZATION IN THE FISHING CAT (PRIONAILURUS VIVERRINUS)

Reproduction Fertility and Development ◽

10.1071/rdv18n2ab284 ◽

2006 ◽

Vol 18 (2) ◽

pp. 249

Author(s):

G. Magarey ◽

J. Herrick ◽

K. Thiangtum ◽

W. Tunwattana ◽

W. Swanson

Keyword(s):

Sperm Motility ◽

Culture Medium ◽

Culture Media ◽

Egg Yolk ◽

Ovarian Follicles ◽

Blastocyst Stage ◽

Motile Sperm ◽

Vitro Fertilization ◽

International Transport

Wild populations of fishing cats (Prionailurus viverrinus) in Southeast Asia are in decline, primarily due to habitat loss. Because the fishing cat population in North American zoos is small (n = 69) and inbred (F = 0.17) with relatively low genetic variation (86%), infusion of new founder genes from Asia is a conservation priority. Importation of cryopreserved semen for use with IVF and ET may offer one alternative to the international transport of living animals. In this study, our objectives were to (1) compare motility longevity of fresh vs. frozen-thawed fishing cat spermatozoa in two culture media, (2) evaluate ovarian responses to exogenous gonadotropins, and (3) assess development of IVF embryos produced with fresh vs. frozen-thawed spermatozoa. Raw semen was collected via electroejaculation from male fishing cats (n = 4), divided into groups, and washed. Two sperm pellets were resuspended in either Ham's F10 medium (HF10; with 5% FBS) or our feline optimized culture medium (FOCM; with 0.4% BSA); another pellet was diluted in TEST egg yolk, cooled to 5�C over 3 h, glycerated (4%), and cryopreserved in straws over LN2 vapor. Frozen sperm samples were thawed, washed, and diluted in either HF10 or FOCM. Fresh and frozen-thawed sperm motility (percent motile, rate of forward progress) in each medium (10 � 106 motile sperm/mL) was assessed (at 0, 1, 3, and 6 h) in microdrops under oil during culture (38�C; 6% CO2 in air). Female fishing cats (n = 10) were treated with exogenous gonadotropins (150 IU eCG, 100 IU hCG, 85-h interval) and ovarian follicles were aspirated laparoscopically. Recovered oocytes were inseminated with fresh (2 � 105 motile sperm/mL) or frozen-thawed (5 � 105 motile sperm/mL) spermatozoa in FOCM microdrops; resulting embryos were either cryopreserved or cultured in FOCM (with 5% FBS added at 72 h post-insemination) for 7 days. Sperm motility over time did not differ (P > 0.05) between media for either fresh or frozen-thawed samples; however, across media, frozen-thawed sperm motility was lower (P < 0.05) and declined faster (P < 0.05) compared to fresh spermatozoa. Females produced an average (�SEM) of 9.8 � 2.9 mature ovarian follicles, allowing recovery of 7.3 � 2.6 high-quality oocytes per female. Oocyte cleavage percentage at 42 h p.i. was lower (P < 0.05) with frozen-thawed spermatozoa (38%, 11/29) compared to freshly collected spermatozoa (68%, 17/25). Overall, 35% (6/17) of cultured embryos developed to blastocysts with no difference (P > 0.05) between embryos produced with frozen-thawed (4/11) vs. fresh (2/6) spermatozoa. Although fishing cat sperm motility and fertility appear compromised after cryopreservation, our results demonstrate the ability of frozen-thawed spermatozoa to produce IVF embryos that are capable of developing to blastocyst stage in vitro. This work was supported by (NIH RR015388).

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Effect of Age and Frequency of Collection in Quality of Jersey Bulls Semen at National Livestock Breeding Center, Nepal

Nepalese Veterinary Journal ◽

10.3126/nvj.v35i0.25213 ◽

2018 ◽

Vol 35 ◽

pp. 9-22

Author(s):

S. Sankhi ◽

K.R. Sapkota ◽

B. Regmi

Keyword(s):

Semen Quality ◽

Sperm Concentration ◽

Quality Parameters ◽

Bull Semen ◽

Livestock Breeding ◽

Abnormal Sperm ◽

Breeding Center ◽

Live Sperm ◽

Level Of Significance ◽

Effect Of Age

The present study was undertaken to determine the effect of age and frequency of collection on quality parameters of Jersey bull semen at National Livestock Breeding Center (NLBC), Pokhara, Kaski district. Nine Jersey bulls were selected randomly from NLBC. To obtain the effect of age, six bulls of three different age interval (3- 4, 5-7 and 8-9 years) were selected randomly. Remaining three bulls were used to determine the effect of frequency of collection on semen quality. Studies were conducted for three months (Nov/Dec to Dec/Jan 2017). Semen quality was analyzed using CASA system and SMILE software. ANOVA and paired t- tests were applied to find out level of significance. All the statistical analyses were done using SPSS 17.0. Sperm concentration (0.551 ±0.004 to 1.084 ± 0.014x109), post- thaw motility (42.44±0.11 to 51.72±0.03 %), live sperm (45.62 ± 0.13 to 51.23±0.15 %), and abnormal sperm (11.57±0.12 to 19.59±0.24%) were significantly affected by the age of the bulls. Semen quality was found superior in the bulls belonging to 5-7 years of age intervals. On the other hand, volume (3.98±0.09 to 6.72±0.06 ml), initial motility (72.68±0.02 to 78.64±0.15 %) and pre-filling motility (71.34±0.17 to 77.62±0.02 %) did not differ significantly among different age intervals. Significantly, higher value for semen volume (3.98±0.12 to 6.02±0.14 ml), sperm concentration (0.572±0.119 to 1.095±0.015) and post- thaw motility (40.34±0.21 to 48.54±0.06% was obtained on second collection. However, initial motility (70.58±0.06 to 75.64±0.13 %), prefilling motility (68.11±0.12 to 71.62±0.02 %), live sperm (53.27±0.54 to 56.17±0.02 %) and abnormal sperm (12.31±0.12 to 14.3±0.47 %) did not differ significantly between the frequencies of collection. Different quality parameters of semen like volume, concentration, initial motility, pre-filling motility, post thaw motility, live and normal spermatozoa percentage were higher in 5-7 years old bulls and in second collection of semen.

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