Search Results (35)

The conservation of small and genetically vulnerable brown bear populations, such as the Cantabrian subpopulation in Spain, depends on developing species-specific assisted reproductive technologies and genetic resource banks. However, the lack of standardized sperm collection and cryopreservation protocols hinders their application. This study provides the first comparative analysis of sperm quality and proteomic profiles from three different origins: epididymal, pre-ejaculated, and ejaculated. Sperm quality parameters —motility and kinetic, viability, apoptosis, and oxidative stress— and protein expression were assessed. Although yields were similar, marked differences were observed in sperm quality and protein profiles. Sixty-three proteins involved in metabolism, stress response, and oxidative balance were differentially expressed depending on sperm origin. Epididymal sperm showed the highest viability and motility, lowest apoptosis, and a proteomic profile indicative of active spermatogenesis and enhanced oxidative stress defense. In contrast, ejaculated sperm had increased oxidative stress and reduced expression of metabolic proteins, while pre-ejaculated sperm exhibited lower motility, likely due to urine contamination and mitochondrial protein alterations, despite comparable viability and apoptosis. These findings offer novel insights into brown bear sperm biology and highlight the importance of sperm origin in developing optimized assisted reproduction strategies, ultimately supporting ex situ conservation efforts for this species. Full article

The Hucul horse is a Polish primitive breed with a small population size, which highlights the importance of preserving the genetic resources. The cryopreservation of semen is essential for creating gene banks, but its effect on the acrosome reaction in Hucul stallions has not yet been investigated. The acrosome reaction is one of the most important physiological events associated with the fertilization process. Therefore, our goal was to determine the level of acrosome reaction in chilled and frozen/thawed Hucul stallion semen using the FluoAcro test and the SCA^® semen analysis system. We found that semen cryopreservation significantly reduced sperm motility and was associated with an increased percentage of acrosome-reacted spermatozoa. It should be noted, however, that, in this case, there was no negative control, and the results may reflect acrosomal damage rather than the elicited responses. Further validation of the methods with equine sperm and the inclusion of a control are recommended. Full article

Objectives: This study aimed to evaluate the implementation of a formalized fertility preservation (FP) program for transgender and nonbinary patients assigned male at birth (TGNB-AMAB) at our institution. Methods: We reviewed TGNB-AMAB patients who were referred to the FP program at our academic institution between 2016 and September 2023. We compared the number of referrals and the percentage of patients who underwent FP per year. Clinical and demographic information including age at referral, time from referral to banking, semen parameters, and serum hormone values were evaluated. Results: In total, 154 TGNB-AMAB patients were referred to the FP program since 2016; 131 (85.1%) met with a reproductive urologist or advanced practice provider for FP consultation; and 124 (94.7%) completed sperm cryopreservation. The number of annual referrals significantly increased over time (p = 0.001). The average age (±standard deviation) at referral was 20.5 ± 5.7 years. The median time from referral to sperm cryopreservation was 14 days. The average semen parameters among all the patients were volume 2.7 ± 1.7 mL, sperm concentration 36.0 ± 31.6 M/mL, sperm motility 56.8 ± 19.0%, and sperm morphology 4.7 ± 2.9%. There was no significant difference in semen parameters between TGNB-AMAB patients previously on gender-affirming hormonal therapy prior to banking and those not on prior hormonal treatment (p > 0.05). Conclusions: Our fertility preservation program significantly increased the number of TGNB-AMAB patients who received consultation and underwent sperm cryopreservation. The institution of a formalized FP program can be used to increase access for TGNB-AMAB patients who desire future fertility. Full article

For adolescents and young adults (AYAs) with cancer, fertility preservation is recommended before starting gonadotoxic treatments. This is an important aspect of psychological support in the treatment of the disease. However, the enormous psychological impact of this procedure on adolescents and young adults with cancer needs to be addressed by professionals. The traumatic nature of cancer diagnosis disrupts the psychosocial development of AYAs. A young adolescent’s perception of reproduction, and in particular of sperm freezing, is greatly altered by the disease. For a teenager, the success of sperm banking results from a positive balance between facilitators and barriers, which are mentioned here. Moreover, this article proposes a symbolic interpretation of sperm banking, referring to landmarks integrated during childhood, especially in fairytales. Furthermore, it offers an original video documentary that can be used as an information support to help AYAs adhere to the process of preserving their fertility through sperm freezing. Full article

This study investigates the efficacy of hormone-induced artificial reproduction in goldfish (Carassius auratus) under controlled temperatures. Ovaprim injections significantly enhanced ovulation and sperm production compared to controls. Medium temperature (22 °C) produced the highest ovulation rates, fastest ovulation timing, and optimal sperm quality (motility and morphology) compared to high (28 °C) and low (16 °C) temperature groups. The low-temperature group exhibited reduced sperm motility duration and higher rates of sperm and larvae damage. The sperm volume of the high-temperature group was higher, but their post-injection survival rates were lower. Furthermore, the lowest spawning rate and low egg quality were noted in the high temperature. Cryopreservation using extender E4 (15% DMSO) exhibited superior post-thaw sperm motility and achieved higher fertilization rates. Fertilization rates, embryo development, and larval survival were all highest at the medium temperature. Larvae hatched from fresh sperm at medium temperature exhibited faster growth and fewer deformities. These findings suggest that hormone stimulation coupled with a medium temperature regimen is critical for successful artificial reproduction in goldfish. Cryopreservation with extender E4 holds promise for sperm banking; however, further optimization is necessary to improve fertilization success with thawed sperm. Future research could explore the influence of temperature on sperm physiology and refine cryopreservation protocols to enhance fertilization rates. Full article

Introduction and Objectives: In the modern context, where fertility is crucial for couples, male factors contribute 40–50% to subfertility. Testicular cancer survivors facing subfertility due to treatments like orchidectomy and chemotherapy can benefit from sperm banking. However, awareness is lacking, especially in culturally sensitive Asian populations where sex and fertility discussions are taboo. This study aims to assess attitudes and utilization of sperm banking, evaluate its impact on pregnancy outcomes, and identify implementation obstacles. Materials and Methods: A phone interview survey targeted testicular cancer patients treated at Hospital Sultanah Aminah Johor Bahru and Sarawak General Hospital in Malaysia (2019–2023). Of the 102 identified patients, 62 participated. Investigators, using contact details from medical records, conducted interviews with a blend of quantitative and qualitative inquiries. Bivariate analysis identified factors linked to the decision to pursue sperm banking. Results: Out of 62 participants, 58.1% were aware of sperm banking, yet 90.3% chose not to utilize it. Reasons for declining included physician non-offer (41.1%), cost concerns (21.4%), a desire for prompt treatment (16.1%), lack of interest (14.3%), and other factors (7.1%). Among six patients opting for sperm banking, 50% utilized banked sperm, resulting in successful progeny for two-thirds. Notably, one case led to multiple pregnancies. Ethnicity (p = 0.046) and religion (p = 0.026) significantly influenced decisions, with Muslim Malays being the least likely to utilize sperm banking. Conclusion: Sperm banking emerges as a cost-effective strategy for safeguarding fertility in testicular cancer patients. Healthcare providers should proactively offer this option before treatment, ensuring patients are well-informed and addressing concerns to foster informed decisions. Full article

Studies revealed a global loss of genetic resources for local sheep breeds. Therefore, the current study aimed to introduce and highlight the progress made on Hungary’s existing gene conservation program (small Gene Bank). Furthermore, we evaluated breed (Tsigai, Cikta, and Racka), season, and individual variabilities (n = 24) of the pre-freeze and post-thaw semen stored in the Gene Bank to enhance the gene conservation of the breeds. The samples were cryopreserved manually, and post-thaw spermatozoa were analyzed for motility (CASA), viability, chromatin structure, and morphometry of the sperm nuclei. Ejaculate volume, spermatozoa concentration, subjective motility and standard motility, kinematic parameters, and spermatozoa’s head area standard deviation of the post-thaw samples differed significantly among breeds (p < 0.05). Season affected ejaculate volume, total spermatozoa number/ejaculate, STR, BCF, and ALH. We observed a significant (p < 0.001; 0.05) breed and season interaction on concentration, total spermatozoa number/ejaculate, VCL, LIN, WOB, spermatozoa’s head average perimeter and nucleus length (Tsigai and Cikta differed but were statistically the same as Racka). Similarly, season significantly (p < 0.05) affected the proportion of ejaculate suitable for freezing. There was a significant (p < 0.05) difference in kinematic parameters and viability among the rams across the breeds. The spermatozoa’s head morphometry of the Tsigai and Cikta breeds differed significantly (p < 0.05) among the rams. There were individual and breed differences in many spermatozoa quality parameters. The stored samples are of good quality, with more than 40% having intact membranes and low abnormal chromatin condensation. Full article

Background: Sperm cryopreservation is recommended to preserve male fertility for cancer patients or other medical conditions at risk of sperm decline. Whether motility and viability recovery rates vary depending on the medical conditions requiring cryopreservation is poorly known. We report here on the 24-year experience of our semen bank. Methods: Motility and viability recovery rates were evaluated in 1973 collections from patients with various medical conditions and 67 collections from donors, and the results were related to basal semen quality. Results: Motility and viability recovery were highly related to basal semen quality and varied between cancer and non-cancer conditions, independently of the duration of cryopreservation and patient age. In samples with a sperm number below 2 × 10⁶/mL, recovery rates approximated to zero. The highest recovery rates were found in donor collections. Cut-off values for the recovery of at least 1% motile spermatozoa were established based on initial semen quality. Conclusions: Our results indicate that the occurrence of any pathological or medical condition resulted in lower recovery rates with respect to donors, indicating that intrinsic sperm characteristics drive susceptibility to cryodamage. Established cut-off values for motility recovery can be useful for patient counseling as well as for ART laboratories to decide the type of procedure. Full article

Semen cryobanks are critical for preserving autochthonous and rare breeds. Since sperm cryopreservation has been optimized for commercial breeds, non-commercial ones (often endangered) must be characterized to ensure the germplasm’s viability. This study reports an investigation of the “Asturiana de la Montaña” breed (AM), a valuable Spanish autochthonous cattle breed adapted to the mountainous Atlantic environment. The survey included cryopreserved semen doses from 40 bulls stored at the Principado de Asturias Germplasm Bank. Data were obtained from the routine fresh semen analysis, CASA (motility), and flow cytometry analyses of fresh and post-thawing semen, and the 56-day non-return-rate (NRR) in heifers and cows (all results as 1st and 3rd quartiles). Fresh samples (artificial vagina) were within the normal range for cattle (4–6 mL, 5–10 × 10⁹/mL; mass motility 5). Post-thawing results showed motility below typical for commercial breeds (total motility 26–43%, progressive 14–28%), with higher values for viability (47–62%). Insemination results showed a good performance for this breed (NRR: 47–56%; higher for heifers). Sperm volume increased with age, with little or no effects on sperm quality. Few associations were found between post-thawing quality or freezability and NRR, LIN being the variable more strongly associated (positively). The AM semen bank shows a good prospect for preserving and disseminating the genetics of this breed. This survey indicates that dedicated research is needed to adapt freezing protocols to this breed, optimizing post-thawing results. Full article

Primary Bladder Neck Obstruction (PBNO) management provides medical and surgical treatment, such as transurethral incisions that can lead to retrograde ejaculation. The aim of this study was to investigate the maintenance of anterograde ejaculation and semen quality before and after this surgical procedure. A retrospective evaluation was carried out between 2011 and 2020. A total of 73 patients diagnosed with PBNO were recruited. Ejaculatory function, semen quality, and the fertility of recruited subjects were evaluated. Semen parameters—Baseline, 8.2% of patients were oligozoospermic and 12.3% had a semen volume below the WHO 2010 fifth percentile. Post-surgery, 20% of patients were oligozoospermic. We detected a significant decrease in total sperm number, a significant increase in the number of abnormal forms, and a reduction in the leukocyte concentration. Ejaculatory function—A total of 7.7% of patients reported anejaculation after transurethral incision of the bladder neck. Fertility—9.2% of the patients already had children before surgery; 13.8% had naturally conceived children in the years following surgery; 76.9% had no desire for paternity at the time. Our data have important implications for sperm bank management. The alterations in semen parameters and the risk of anejaculation suggest that the use of sperm cryopreservation before surgery for PBNO should be encouraged. Full article

Cryopreservation is an expanding strategy to allow not only fertility preservation for individuals who need such procedures because of gonadotoxic treatments, active duty in dangerous occupations or social reasons and gamete donation for couples where conception is denied, but also for animal breeding and preservation of endangered animal species. Despite the improvement in semen cryopreservation techniques and the worldwide expansion of semen banks, damage to spermatozoa and the consequent impairment of its functions still remain unsolved problems, conditioning the choice of the technique in assisted reproduction procedures. Although many studies have attempted to find solutions to limit sperm damage following cryopreservation and identify possible markers of damage susceptibility, active research in this field is still required in order to optimize the process. Here, we review the available evidence regarding structural, molecular and functional damage occurring in cryopreserved human spermatozoa and the possible strategies to prevent it and optimize the procedures. Finally, we review the results on assisted reproduction technique (ARTs) outcomes following the use of cryopreserved spermatozoa. Full article

Purpose: To evaluate the effectiveness of donor in vitro fertilization (IVF-D) and donor artificial insemination (AI-D) in clinical outcomes, risks, and costs. Methods: This study analyzed the cycle changes and clinical outcomes in 20,910 IVF-D and 16,850 AI-D cycles between 2013 and 2021 in the Reproductive and Genetic Hospital of CITIC-Xiangya. A cost-effectiveness analysis was performed to evaluate the costs per couple and per live birth cycle in the two treatment groups. Results: IVF-D had higher pregnancy and live birth rates than AI-D (p < 0.001). The cumulative pregnancy and live birth rates for three AI-D cycles were 41.01% and 32.42%, respectively, higher than the rates for one or two AI-D cycles. The multiple birth and birth defect rate of AI-D was lower than that of IVF-D significantly. IVF-D mean cost per couple was higher than that of AI-D (CNY32,575 vs. CNY11,062, p < 0.001), with a mean cost difference of CNY21,513 (95% confidence interval, CNY20,517–22,508). The mean costs per live birth cycle for IVF-D and AI-D were CNY49,411 and CNY31,246, respectively. Conclusion: AI-D is more cost-effective and poses a lower risk for infertility couples than IVF-D, and patients should undergo three AI-D cycles to obtain the highest success rate. Full article

Sperm cryopreservation and biobanking are emerging as tools for supporting genetic management of small and threatened populations in amphibian conservation programs. However, there is little to no evidence demonstrating reproductive maturity and viability of offspring generated with cryopreserved sperm, potentially limiting widespread integration of these technologies. The purpose of this report is to demonstrate that amphibian sperm can be cryopreserved and thawed to successfully produce individuals of an F1 generation that can reach adulthood and reproductive maturity, to generating viable gametes and an F2 generation. Species-specific exogenous hormones were administered to both F0 and F1 adults to stimulate spermiation and oviposition in the eastern tiger salamander (Ambystoma tigrinum), dusky gopher frog (Lithobates sevosa), and Puerto Rican crested toad (Peltophryne lemur). Sperm cells collected non-lethally from F0 adults were cryopreserved, thawed, and used for in vitro fertilization (IVF) to produce F1 offspring. Individuals of the F1 generation are shown to reach adulthood, express viable gametes, and produce offspring through facilitated breeding, or IVF. The production of amphibian F2 generations shown here demonstrates that amphibian sperm collected non-lethally can be banked and used to generate reproductively viable animals of subsequent generations, thus maintaining valuable genetic linages and diversity in threatened amphibian species. The incredible value that cryopreservation of sperm has for long-term genetic management aids in the sustainability of both in situ and ex situ conservation efforts for this taxon. Full article

Cryopreservation is a way to preserve germplasm with applications in agriculture, biotechnology, and conservation of endangered animals. Cryopreservation has been available for over a century, yet, using current methods, only around 50% of spermatozoa retain their viability after cryopreservation. This loss is associated with damage to different sperm components including the plasma membrane, nucleus, mitochondria, proteins, mRNAs, and microRNAs. To mitigate this damage, conventional strategies use chemical additives that include classical cryoprotectants such as glycerol, as well as antioxidants, fatty acids, sugars, amino acids, and membrane stabilizers. However, clearly current protocols do not prevent all damage. This may be due to the imperfect function of antioxidants and the probable conversion of media components to more toxic forms during cryopreservation. Full article

Thawing protocols have been barely studied, and their modifications may lead to a substantial improvement in post-thawing sperm quality, which could be of great relevance to existing sperm banks, such as those for Iberian pig breeds with varieties in danger of extinction. For that, the study aimed to evaluate different thawing rates and to evaluate modifications in the composition of the thawing extender (basic pH to 8–8.2, incorporation of cyclodextrins loaded with cholesterol [CLC] and the incorporation of ion chelators [EDTA and EGTA]). After thawing, overall sperm motility and kinematic parameters, acrosome status and sperm membrane integrity were evaluated. The most optimal results were obtained with the thawing rate reaching 70 °C for 8 s with the inclusion of 12.5 mg of CLC/500 × 10⁶ spermatozoa in the thawing extender, which showed an improvement compared to the control at 70 °C. In conclusion, to adapt the thawing conditions may be relevant, especially for endangered species or breeds such as some varieties of Iberian pig, since this process could also be used in samples cryopreserved in gene banks. Full article