scholarly journals First Evaluations and Cryopreservation of Semen Samples from Sunda Clouded Leopards (Neofelis diardi)

Animals ◽  
2020 ◽  
Vol 10 (6) ◽  
pp. 1072 ◽  
Author(s):  
Zainal Zainuddin ◽  
Mohamed Mohamed Tarmizi ◽  
Keng Yap ◽  
Pierre Comizzoli ◽  
Symphorosa Sipangkui

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.

2017 ◽  
pp. 99-104
Author(s):  
V.V. Orlova ◽  
◽  
L.V. Suslikova ◽  

The prevalence of infertility, including tubal-peritoneal genesis, led to the development of assisted reproductive technology (ART) as the most effective treatment and achievement of the expected pregnancy. But the success of ART is about 40% and depends on many factors: genetic, immunological, hormonal, age, control ovarial stimulation, sperm quality, quantity and quality received oocytes and embryos and implantation ability of endometrium. Therefore, it is important to consider the mechanisms and factors behind the successful implantation. Particular attention is given to the study of disturbances of oxidative stress and apoptosis in the reproductive tract of infertile women. Under the influence of adverse factors and in pathology, as a result of reducing the cell’s ability to neutralize free radicals and active forms of oxygen, antioxidant protection is disturbed, and oxidative stress develops, which promotes the onset of apoptosis. The programmed cell death occurs in the physiological functioning of the reproductive system: follicular atresia and regression of the functional layer of the endometrium when pregnancy is not occur. However, today the negative influence of oxidative stress and violation of the regulation of apoptosis on the reproductive function, namely the success of in vitro fertilization, has been confirmed. Also, the use of ART has some of the negative effects on gametes and embryos, which is accompanied by the emergence of oxidative stress and reduces the probability of pregnancy. Melatonin is a universal hormone that plays a key role in the functioning of sex hormones and has powerful antioxidant properties. As a key antioxidant regulator As a key antioxidant regulator at the biochemical and hormonal levels, melatonin positively influences the maturation of oocytes and the preparation of the endometrium before implantation. Therefore, there is a reasonable justification for the use of melatonin during treatment with ART. Many studies are devoted to the study of the effectiveness of the use of exogenous melatonin in the treatment of infertility, but to date, the level of evidence is insufficient for use in it ART. Key words: melatonin, infertility, oxidative stress, apoptosis, ART.


2005 ◽  
Vol 17 (2) ◽  
pp. 276 ◽  
Author(s):  
J. Pryor ◽  
S. Romo ◽  
D.D. Varner ◽  
K. Hinrichs ◽  
C.R. Looney

In commercial bovine in vitro fertilization (IVF) companies, there is a continuous need to improve results. Efforts to maximize in vitro embryo production have included modifications in the use of sperm separation gradients. The development of commercially available sperm centrifugation gradients represents a new possibility of increasing the number of viable sperm that can be obtained from low concentration (fresh or frozen, sexed or unsexed) semen samples in order to improve the efficiency of the IVF system to make embryo production as efficient as possible. The objective of this study was to compare two different separation gradients, as follows: Group 1: Percoll (Sigma, St. Louis, MO, USA), in 45% and 90% gradients; Group 2: EquiPure (Nidacon, Gathenburg, Sweden), in top and bottom layers. Before and after separation, sperm were evaluated at 200× magnification for total motility, and then stained to assess viability at 400× with fast-green/eosin stain (Sigma). Sperm separation was performed using frozen/thawed semen from one bull. Semen was separated by centrifugation at 200g for 30 min in both density gradients. Results obtained from Groups 1 and 2 were compared by chi-square test. Sperm separation with Percoll yielded lower numbers of sperm (average sperm concentration after separation of 92 × 106, vs. 159 × 106 sperm/mL for EquiPure; P < 0.05) but resulted in higher motility (60% vs. 39%, respectively; P < 0.05) of separated sperm. Rates of live sperm cells were not significantly different between groups (69.5% vs. 70%, respectively; P > 0.1). These results indicate that the commercial separation medium EquiPure may be associated with higher sperm concentration levels but with lowered sperm motility when compared to Percoll for bovine sperm separation. However, Equipure provided similar percentages of live sperm when compared to Percoll, which is currently used in our laboratory.


2019 ◽  
Vol 55 ◽  
pp. 58-61 ◽  
Author(s):  
Hanne Skovsgaard Pedersen ◽  
Ying Liu ◽  
Leslie Foldager ◽  
Henrik Callesen ◽  
Knud Larsen ◽  
...  

2020 ◽  
Vol 3 (2) ◽  
pp. 99-106
Author(s):  
Sara Mahmood Qureshi ◽  
Salma Kafeel ◽  
Riffat Bibi ◽  
Jawad Mohmand

Introduction: The unrestricted use of intracytoplasmic sperm injection (ICSI) for non-male factor infertility is associated with adverse outcomes. Post-wash total motile sperm count (PW-TMSC) offers prognostic value to assess sperm quality and aid in the decision to perform in vitro fertilization (IVF) or ICSI. Objectives: The aim of this study was to identify the effect of PW-TMSC on fertilization rates in patients undergoing IVF cycles exclusively with non-male factor infertility. It also aimed to identify whether unnecessary ICSI could be avoided in such cases, thus maximizing optimal outcomes. Materials & Methods: We retrospectively analyzed age, semen volume, prewash TMSC, and PW-TMSC in 68 conventional IVF cycles of infertile couples with non-male factor infertility. Clinical characteristics including female age, number of follicles, level of estradiol on trigger day, mature cumulus-oocyte complexes (COCs) collected, were also included. Results: Incidence of <30% fertilization was significantly higher in the 4-<10 Million group compared with the ≥20 Million post-wash TMSC group (P<0.001). Furthermore, Receiver operating characteristics (ROC) analysis revealed post-wash TMSC as a significant predictor (P<0.05) of total failed fertilization (TFF) and of ≥30% fertilization (P<0.05) with area under curve (AUC) of 0. 79 and 0.77, respectively, with a deemed cutoff of 10.89 Million. Conclusion: Post-wash TMSC is a good predictor of fertilization; it can help in avoiding potentially low or even total fertilization failure (TFF). A cut-off point of 10.89 Million or less should warrant the use of ICSI.


Author(s):  
Herman J. Tournaye

Anamnesis, physical examination, and additional tests may reveal a specific cause of reproductive failure in infertile men. Whenever this is found, a specific treatment or cure should be applied. When no such treatment is available, or when specific treatment has failed, techniques of assisted reproduction may be proposed to couples suffering from long-standing male infertility. The rationale behind these is to bring the spermatozoa closer to the oocyte in an attempt to enhance the fertilization process. In recent years the role of assisted reproduction has become more important, and it has often been stated that these techniques have made clinical work-up or specific treatment of the male partner pointless. However, this is far from true. Not only may correction of a specific dysfunction in the male avoid the use of assisted reproductive techniques, but careful work-up and treatment may also enhance the outcome of these treatments. Assisted reproductive techniques should not be viewed as a primary treatment option, but rather as a complementary treatment when other treatments have failed, or have been judged inadequate after a complete work-up.


2019 ◽  
Vol 210 ◽  
pp. 106178 ◽  
Author(s):  
Zhiqiang Ren ◽  
Weike Shaoyong ◽  
Qian Li ◽  
Lu Ma ◽  
Junying Xiao ◽  
...  

eLife ◽  
2017 ◽  
Vol 6 ◽  
Author(s):  
Jean-Ju Chung ◽  
Kiyoshi Miki ◽  
Doory Kim ◽  
Sang-Hee Shim ◽  
Huanan F Shi ◽  
...  

We report that the Gm7068 (CatSpere) and Tex40 (CatSperz) genes encode novel subunits of a 9-subunit CatSper ion channel complex. Targeted disruption of CatSperz reduces CatSper current and sperm rheotactic efficiency in mice, resulting in severe male subfertility. Normally distributed in linear quadrilateral nanodomains along the flagellum, the complex lacking CatSperζ is disrupted at ~0.8 μm intervals along the flagellum. This disruption renders the proximal flagellum inflexible and alters the 3D flagellar envelope, thus preventing sperm from reorienting against fluid flow in vitro and efficiently migrating in vivo. Ejaculated CatSperz-null sperm cells retrieved from the mated female uterus partially rescue in vitro fertilization (IVF) that failed with epididymal spermatozoa alone. Human CatSperε is quadrilaterally arranged along the flagella, similar to the CatSper complex in mouse sperm. We speculate that the newly identified CatSperζ subunit is a late evolutionary adaptation to maximize fertilization inside the mammalian female reproductive tract.


2005 ◽  
Vol 17 (2) ◽  
pp. 277
Author(s):  
S. Romo ◽  
J. Pryor ◽  
D.D. Varner ◽  
K. Hinrichs ◽  
C.R. Looney

Recently, the development of commercially available defined media and sperm centrifugation gradients has offered new possibilities for increasing the efficiency of commercial in vitro fertilization (IVF) systems. The objective of this study was to compare three different IVF protocols using two different separation gradients, two fertilization media, and two embryo culture media, as follows: Group 1. sperm separation (SS): Percoll (Sigma, St. Louis, MO, USA), fertilization medium (FM): TALP-Fert (TFM), embryo culture media (ECM): G1/G2 (version 3, Vitrolife, Englewood, CO, USA). Group 2. SS: Percoll, FM: Bovine vitro Fert (Cook, Brisbane, Australia), ECM: Bovine vitro Blast/Bovine vitro Cleave (Cook); and Group 3. SS: EquiPure (Nidacon, Spectrum Technologies, Healdsburg, CA, USA), FM: TFM, ECM: G1/G2. Oocytes were obtained from slaughterhouse ovaries and matured in vitro (Looney et al. 1994 Theriogenology 41, 67). IVF was conducted using frozen/thawed semen from one bull. Semen was separated by centrifugation at 700g for 30 min in the given density gradients; Percoll was used in a 45% to 90% gradient. Sperm viability after separation was assessed by fast-green/eosin stain (Sigma). IVF was carried out in 0.5 mL of the given fertilization medium supplemented with PHE1 and heparin (10 μg/mL), in humidified 5% CO2 in air atmosphere at 38.7°C. Final sperm concentration in the IVF wells was 1 × 106/mL. In Experiment 1, a total of 368 oocytes (2 replicates) were fixed and stained (Hoechst 33342, Sigma) 24 h post-IVF to assess sperm penetration (Group 1, n = 128, Group 2, n = 108, Group 3, n = 132). In Experiment 2, a total of 400 embryos (2 replicates) were cultured in 0.5 mL of the given culture medium under mineral oil in a 5% O2, 5% CO2, 90% N2 atmosphere at 38.7°C with high humidity for 112 h before fixation and staining. Embryos in Groups 1 (n = 129) and 3 (n = 139) and Group 2 (n = 132) were changed to G2 and Cleave media, respectively, at 84 h. Sperm separation with Percoll yielded lower numbers of sperm (average sperm concentration after separation of 218 vs. 383 × 106 for EquiPure; P < 0.05), but resulted in higher total motility (60% vs. 41%, respectively; P < 0.05) and higher viability (93% vs. 70%, respectively; P < 0.05) of separated sperm. In Experiment 1, rates of normal fertilization were significantly lower for Group 3 (58%) than for Groups 1 and 2 (74% and 77%, respectively, P < 0.05). In Experiment 2, rates of development to <8, 9 to 16, and >16 cells at 112 h were not significantly different among groups (43, 48, and 46% for Group 1; 22, 18, and 31% for Group 2; and 35, 34, and 23% for Group 3, respectively; P > 0.1). These results indicate that the commercial separation medium, EquiPure, may be associated with lowered sperm motility, viability, and fertilization rates when compared to a standard medium (Percoll) for bovine sperm separation. Commercial fertilization and embryo culture media (Bovine vitro Fert, Cleave, and Blast) provided equivalent embryo development to that currently in use by our laboratory (TFM, G1/G2).


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