Development Of Assisted Reproductive Technologies Research Articles

Genomic insights into southern white rhinoceros (Ceratotherium simum simum) reproduction: Revealing granulosa cell gene expression

In vivo-collected granulosa cells (GC) from the southern white rhinoceros (SWR) provide a non-invasive assessment of the developmental status of oocytes prior to in vitro culture, which could aid in the development of assisted reproductive technologies (ART). Our study aimed to investigate gene expression in SWR granulosa cells, collected in vivo and gain preliminary insight into the transcriptional activity occurring within the cells during various stages of oocyte development. It was hypothesized there would be similarities between the SWR GC transcriptome and cattle and humans, two species for which well-annotated genomes are available and ART are commonly used. GC were collected from SWR following ovum pickup (OPU) and pooled from all aspirated follicles. Total RNA was isolated, libraries prepared, and sequencing performed using an Illumina NextSeq 500. Reads were aligned and annotated to CerSimCot1.0. Databases for cattle and human were acquired for comparison. This study identified 37,407 transcripts present in GC of SWR. It was determined that cattle and human transcriptomes are valuable resources with a homology of 45 % with the SWR. In conclusion, these data provide preliminary, novel insights into the transcriptional activity of GC in the SWR that can be used to enhance ART in this species.

Succinate dehydrogenase participation in porcine gamete function

Context Porcine gametes require energy for the physiological processes that allow fertilisation. Succinate dehydrogenase (SDH) plays a pivotal role in both, the tricarboxylic acid (TCA) cycle and the respiratory chain. Aims The aim of this work was to study the participation of SDH in the in vitro oocyte maturation, sperm capacitation and acrosome reaction in porcine species. Methods Cumulus–oocyte complexes (COCs) from abattoir-derived porcine ovaries were collected by aspiration and were incubated in maturation media, with the addition of increasing concentrations (0, 1, 5 and 10 mM) of malonate (a specific inhibitor of SDH). Nuclear maturation and cytoplasmatic maturation were analysed. Semen samples were incubated for 2 h in capacitating medium with 40 mM sodium bicarbonate, as sperm capacitation inducer, and the addition of increasing concentrations of malonate (0, 1, 5 and 10 mM). Sperm capacitation state and true acrosomal reaction were evaluated. SDH activity was determined in sperm and oocyte extracts by the spectrophotometric method. Key results The addition of 10 mM of malonate decreased both nuclear and cytoplasmic maturation rates (P < 0.05) without affecting COC viability (assessed using fluorescein diacetate). A lower level of capacitation (induced by bicarbonate) and acrosome reaction (induced by follicular fluid) was observed with the addition of 5 mM of malonate (P < 0.05) without affecting motility and viability of sperm at this concentration. The activity of SDH was 0.35 ± 0.1 × 10−5 and 2.37 ± 0.9 × 10−5 U/COC for immature and in vitro matured COC extracts (P < 0.05) respectively, and 0.44 ± 0.16 U/1010 sperm for boar sperm extracts. Conclusions In conclusion, because it has been proposed that aerobic and anaerobic metabolic pathways of cells are changed depending on the oxygen availability and the composition of metabolic substrates in their environment, our results suggest that energy obtained through the mitochondrial respiration (TCA cycle and oxidative phosphorylation) is necessary to support oocyte maturation, sperm capacitation and acrosome reaction in the porcine species. Implications The study of enzymatic activity in gametes is essential for understanding the mechanisms that control the energy production required to achieve successful fertilisation. This knowledge has significant implications for the development of assisted reproductive technologies.

The Umbilical Cord and Complications of Twin Gestations

Abstract The rate of twin pregnancies has increased over the last decades, largely because of the ongoing development of assisted reproductive technology and increased maternal age at childbearing. Twins have a higher risk of adverse outcomes during pregnancy and the perinatal period. The prevalence of umbilical cord abnormalities is higher for twin pregnancies compared with singleton pregnancies. Some of these abnormalities are nonspecific to twinning and can also be found in singleton gestations (such as velamentous cord insertion, vasa previa, and single umbilical artery). Other abnormalities are associated with monochorionic twins, such as umbilical cord entanglement, and umbilical proximate cord insertion. Most of these abnormalities can be detected by ultrasound evaluation. The early and accurate ultrasound diagnosis of chorionicity, amnionicity, and placental and umbilical cord characteristics is crucial if we are to predict the risk of complications and to determine the best management for twin pregnancies. Histopathological examination of the placenta and umbilical cord after delivery can help to confirm prenatal diagnosis and to provide a better understanding of the physiopathology of their abnormalities. The aim of this review was to emphasize the role that the umbilical cord plays in twin complications and to describe the management of these high-risk pregnancies.

Gene mutations impede oocyte maturation, fertilization, and early embryonic development.

Reproductive diseases are a long-standing problem and have become more common in the world. Currently, 15% of the world's population suffers from infertility, and half of them are women. Maturation of oocytes, successful fertilization, and high-quality embryos are prerequisites for pregnancy. With the development of assisted reproductive technology and advanced genetic assays, we have found that infertility in many young female patients is caused by mutations in various developmental regulators. These pathogenic factors may result in impediment of oocyte maturation, failure of fertilization or early embryonic development arrest. In this review, we categorize these clinically-identified, mutated genetic factors by their molecular characteristics: nuclear factors (PALT2, TRIP13, WEE2, TBPL2, REC114, MEI1 and CDC20), cytoplasmic factors (TLE6, PADI6, NLRP2/5, FBXO43, MOS and BTG4), a factor unique to primates (TUBB8), cell membrane factor (PANX1), and zona pellucida factors (ZP1-3). We compared discrepancies observed in phenotypes between human and mouse models to provide clues for clinical diagnosis and treatment of related reproductive diseases.

Female sterilization reversal in the era of in-vitro fertilization.

Regret after female sterilization is not uncommon in the United States. Prior to the development of assisted reproductive technology (ART), surgical reversal of sterilization was the only option for patients interested in fertility. First performed in 1972, this procedure has since been refined over the years by gynaecologic surgeons. With in-vitro fertilization (IVF) gaining popularity, interest in sterilization reversal has waned. However, sterilization reversal should remain an important option in patients seeking pregnancy after tubal ligation. A direct comparison between IVF and sterilization reversal is challenging due to inherent differences in reporting fertility outcomes. However, sterilization reversal may optimize fertility in younger women, whereas IVF may be more effective in older women. The surgical approach to sterilization reversal can be laparotomic, laparoscopic or robotic. Clinical decision making should include consideration of the risk of ectopic pregnancy, interval from sterilization to reversal, type of sterilization procedure, planned anastomotic site and projected remaining tubal length. In the era of IVF, sterilization reversal still has a place in the management in restoring fertility. Creating awareness of the role of sterilization reversal is the first step in improving access to adequate training in this procedure for the next generation of reproductive surgeons.

Temporal trends and adverse perinatal outcomes of twin pregnancies at differing gestational ages: an observational study from China between 2012–2020

BackgroundWith the development of assisted reproductive technology, the twinning rate in China has been increasing. However, little is known about twinning from 2014 onwards. In addition, previous studies analysing optimal gestational times have rarely considered maternal health conditions. Therefore, whether maternal health conditions affect the optimal gestational time remains unclear.MethodsData of women delivered between January 2012 and December 2020 were collected through China’s National Maternal Near Miss Surveillance System. Interrupted time series analysis was used to determine the rates of twinning, stillbirth, smaller than gestational age (SGA), and low Apgar scores (< 4) among twins in China. To estimate the risk of each adverse perinatal outcome for separate gestational weeks, a multivariate generalised linear model was used. Infants born at 37 weeks of gestational age or foetuses staying in utero were used as reference separately. The analyses were adjusted for the sampling distribution of the population and the cluster effect at the hospital and individual levels were considered.ResultsThere were 442,268 infants enrolled in this study, and the adjusted rates for twinning, stillbirth, SGA, and low Apgar scores were 3.10%, 1.75%, 7.70%, and 0.79%, respectively. From 2012 to 2020, the twinning rate showed an increasing trend. Adverse perinatal outcomes, including stillbirth, SGA, and low Apgar scores showed a decreasing trend. A gestational age between 34 and 36 weeks decreased most for rate of stillbirth (average changing rate -9.72%, 95% confidence interval [CI] -11.41% to -8.00%); and a gestational age of between 37 and 38 weeks decreased most for rates of SGA (average changing rate -4.64%, 95% CI -5.42% to -3.85%) and low Apgar scores (average changing rate -17.61%, 95% CI -21.73% to -13.26%). No significant difference in changes in twinning rate or changes of each perinatal outcome was observed during periods of different fertility policies. Infants born at 37 weeks of gestation had a decreased risk of stillbirth, SGA, and low Apgar scores. Maternal antepartum or medical complications increased the risk of SGA and low Apgar scores in different gestational weeks.ConclusionChina’s twinning rate showed an increasing trend, while adverse perinatal outcomes decreased from 2012 to 2020. Fertility policy changes have had little effect on the twinning rate or the rate of adverse perinatal outcomes such as stillbirth, SGA, or low Apgar scores. The optimal gestational age for twins was 37 weeks. Women pregnant with twins and with antepartum or medical complications should be cautious due to an increased risk of SGA and low Apgar scores.

Correlation analysis of adverse outcomes for the selective reduction of twin pregnancies

BackgroundDue to the extensive development of assisted reproductive technology, the number of twin pregnancies has increased significantly over recent decades. Twin pregnancy is the most representative type of multiple pregnancies and is associated with high infant morbidity and mortality. Perinatal complications of twin pregnancy are also markedly increased compared with those of single pregnancy. Transabdominal selective reduction (SR) is a remedial intervention. This study aimed to research the adverse outcomes of transabdominal selective reduction of twin pregnancy and the correlation between the reduction week and pregnancy outcomes.ObjectiveThe purpose of this study was to examine the adverse outcomes of the transabdominal selective reduction of twin pregnancy and the correlation between the reduction week and pregnancy outcomes.MethodsA retrospective cohort study of the transabdominal reduction of twin pregnancy was conducted in a single prenatal diagnosis medical centre from September 2012 to October 2020. According to chorionicity, women with twin pregnancies were divided into 2 groups: dichorionic (DC) twin pregnancies and monochorionic (MC) twin pregnancies. Women with DC twin pregnancies underwent potassium chloride reduction, and those with MC twin pregnancies underwent radiofrequency ablation (RFA). The reduction indications included pregnancy complications, foetal abnormalities, and maternal factors. The perinatal outcomes of different chorionic twins after reduction were analysed. Each foetus with an adverse outcome was included. The relative relationship between the reduction weeks and delivery weeks of twins was examined by correlation analysis.ResultsA total of 161 women were included in this study. A total of 112 women had DC twin pregnancies, and 49 women had MC twin pregnancies. Preterm delivery rates were significantly higher in the MC twin reduction group than in the DC twin reduction group prior to 37 weeks (53.1% vs. 29.5%, P = 0.004). The mean gestational age at delivery of the foetuses in the DC twin group that underwent SR was significantly older than that of those in the MC twin group that underwent SR (36.9 ± 4.0 vs. 33.5 ± 6.6 weeks, P = 0.001). The number of DC twins that underwent SR and were delivered after 37 weeks was obviously greater than that of the MC twins that underwent SR (70.5% vs. 46.9%, P = 0.004). The foetal survival rate was 95.5% in the DC twin reduction group and 77.6% in the MC twin reduction group. If the indication of TTTS was not included, there was no significant difference in the foetal survival rate of the DC and MC twin reduction groups (95.5% vs. 86.2%, P = 0.160). Cotwin death 1 week after reduction was greater in the MC group (6.1% vs. 0%, P = 0.027). Compared to other indications, this finding indicated that a significantly lower proportion of women remained undelivered after selective reduction with the indication of TTTS. There was a significant negative correlation between the reduction weeks and delivery weeks of the two groups (P < 0.01), and the best opportunity for reduction was before 22 weeks of gestation.ConclusionThese findings highlighted an obviously negative correlation between the reduction week and delivery week. The transabdominal selective reduction of twin pregnancy should be considered for a lower rate of miscarriage or premature delivery if the reduction week takes place earlier in pregnancy. The rate of preterm delivery was the lowest when transabdominal selective reduction was completed before 22 weeks of gestation. Compared with other RFA indications, a higher rate of premature delivery was shown for MC twins with a reduction indication of TTTS. TTTS with sIUGR might be one of the reasons for the adverse outcomes of reduction for MC twin pregnancy.

Mammalian sperm capacitation: In vivo and in vitro juxtaposition

The development of assisted reproductive technologies (ART) in mammalian species such as in vitro embryo production (IVEP) has the potential to provide great benefits for significant population increase, improve genetic performance and advancement, and reduce transmission of venereal diseases. Correspondingly, in vitro capacitation of sperm is also paramount, related to the ability of sperm to fertilize oocytes, and was created to imitate in vivo conditions in the female reproductive tract. Amid in vitro capacitation developments, studies on how far in vitro capacitation has progressed in mimicking in vivo scenes have not been thoroughly reviewed as a comparative form. Therefore, the present study outlined the series of alterations in mammalian sperm capacitation during their journey in the female reproductive tract by exploring and juxtaposing processes under in vivo and in vitro conditions. Several essential aspects that become gaps between in vivo and in vitro were also identified and elaborated comprehensively in this systematic literature review. We noted that although in vitro capacitation procedures in certain mammalian species have made promising progress and improvements, it is still poorly successful in other species like horses. Our findings further postulated that the occurrence of cryocapacitation, the high ratio of capacitated sperm/oocyte required for successful fertilization, and the incidence of polyspermy cause capacitation under in vitro settings is less efficient and not yet fully comparable to in vivo. This work is therefore proposed several aspects that need to be bettered from in vitro milieu to make it analogous to in vivo environments in modulating sperm capacitation.

СТАНОВЛЕНИЕ И ОРГАНИЗАЦИЯ РЕПРОДУКТОЛОГИИ В РЕСПУБЛИКЕ БАШКОРТОСТАН

Significance. The history of development of assisted reproductive technologies (ART) in the world is detailed in publications. The history of evolvement of reproductology in Russia, especially in the regions, is understudied, substantiating relevance of the study The purpose of the study. To analyze and summarize material on evolvement and organization of reproductology in the Republic of Bashkortostan. Material and methods. Analysis of health data of the Republic of Bashkortostan, data of official websites of medical organizations. Results. In the Republic of Bashkortostan, in 2006, for the first time ever, the ART department was opened at the Republican Perinatal Center, 25 procedures were carried out at the expense of the republican budget. Since 2013, in-vitro fertilization (IVF) has been introduced into the compulsory health insurance program. Currently, 5 medical centers in the Republic take part in the program of state guarantees with the use of ART methods. Conclusion. The study results demonstrate stages and successes of reproductology in the Republic for 15 years; effectiveness of IVF programs exceeds the Russian average.

Legal Identification of Parent-Child Relationship between Lesbian Partners From the Perspective of Using Human Assisted Reproductive Technology to Procreate Children

With the innovation and development of assisted reproductive technology, it is possible for same-sex partners to have their children. More and more same-sex partners choose to have their children through assisted reproductive technology. However, China’s legislation on assisted reproductive technology is not perfect, and the reproductive rights of same-sex partners cannot be guaranteed. This paper selects the related issues of lesbian partners with assisted reproductive technology to have children, through literature research, case analysis, comparative research and other methods, based on the legitimacy of the reproductive rights of lesbian partners, to explore the feasibility of parent-child relationship legal identification of lesbian partners. In order to protect the interests of lesbian partners and their children born through human assisted reproductive technology, the corresponding legal recognition model of parent-child relationship should be constructed.

Вплив антифосфоліпідних антитіл на ризик невдалої імплантації після запліднення in vitro

Infertility is a unique pathology because it concerns a couple, not one person, and is the cause of a whole range of medical, social, demographic and economic problems. With the active development of assisted reproductive technologies, the demand for solving the problem of infertility has grown significantly. However, the prevalence of infertility remains stable, indicating many «white» spots in the diagnosis and treatment of this pathology and determining the search for new risk factors, diagnostic criteria, and treatment algorithms. Purpose — to analyze the factors that may affect implantation, namely the presence of lupus anticoagulant and antiphospholipid antibodies in women with infertility who have undergone in vitro fertilization. Materials and methods. A survey of 106 women diagnosed with infertility underwent treatment with assisted reproductive technologies in September 2019 — February 2021. The control (1) group consisted of women who received infertility treatment according to the standard scheme: controlled ovarian stimulation, gamete collection, intracytoplasmic sperm injection and embryo transfer. The main (2) group included patients who received aspirin, anticoagulants, and intravenous IgG immunoglobulins in addition to the standard treatment regimen. In the present study, levels of lupus anticoagulant and antiphospholipid antibodies were determined in both groups. Results. The study found no differences between groups of patients in levels of antibodies (AB) to phospholipids (IgG), beta-2-glycoprotein (IgG, IgM) and lupus anticoagulant (screening, confirmation) in the two groups of patients (p&gt;0.05). Difference in two groups were in levels of AB to beta-2-glycoprotein IgM in controls, on average, 9.42 units/ml (5.86 units/ml — 12.35 units/ml), and in patients of main group — 7.23 units/ml (3.14 units/ml — 10.58 units/ml), p=0.015. One-factor logistic regression models were used to clinically understand the influence of factors associated with the risk of implantation failure. The analysis did not reveal a connection between risk of implantation failure and levels of autoantibodies (aAB) to phospholipids (IgG, IgM), beta-2-glycoprotein (IgG, IgM) and lupus anticoagulant (screening, confirmation) in two groups of patients (p&gt;0.05 in all cases). Conclusions. The presence of aAB and/or antiphospholipid syndrome during the establishment of infertility factors is an essential component of the algorithm for diagnostic search for the cause of infertility and miscarriage. However, based on the obtained data, the presence of aAB in the study does not affect the implantation process nor affects the treatment of infertility. The research was carried out in accordance with the principles of the Helsinki declaration. The study protocol was approved by the Local Ethics Committee of the participating institution. The informed consent of the patient was obtained for conducting the studies. No conflict of interest was declared by the authors. Key words: infertility, implantation failure, antiphospholipid syndrome, assisted reproductive technologies.

Development of assisted reproductive technologies for the conservation of Atelopus sp. (spumarius complex)

Amphibians are in peril, given the ongoing sixth mass extinction of wildlife. Thus, Conservation Breeding Programs (CBPs) are attempting to breed some species under laboratory conditions. The incorporation of assisted reproduction technologies (ARTs), such as hormonal stimulation, sperm collection and cryopreservation, and in vitro fertilization is contributing to successful CBPs. The objective of this study was to apply ARTs in sexually mature individuals of an undescribed species of Atelopus (spumarius complex) (harlequin frog). Our procedure involves hormonal induction of gametogenesis in this species. We were able to induce gamete release through administration of human chorionic gonadotropin (hCG) in males, and in females this has been achieved through the sequential administration of hCG (priming doses), and combinations of hCG with gonadotropin releasing hormone analogue, GnRHa (ovulary dose). We standardized sperm cryopreservation by performing toxicity tests of cryoprotectants, fast/slow freezing and thawing, as well as supplementation of non-penetrating cryoprotectants (sugars). Next, we performed in vitro fertilization, evaluated the fertilization capacity of the cryopreserved sperm, and describe external features of fresh and cryopreserved sperm. We found that 10 IU/g hCG induced the release of the highest sperm concentrations between 3 and 5 h post-injection, while 2.5 IU/g hCG induced the release of eggs in most treated females. Under cryopreservation conditions, the highest recovery of forward progressive motility or FPM was 26.3 ± 3.5%, which was obtained in cryosuspensions prepared with the 5% DMF and 2.5% sucrose. Cryopreserved sperm showed narrower mitochondrial vesicles after thawing, while in frozen samples without cryodiluent showed 31% of spermatozoa lost their tails. In most cases, our attempts of in vitro fertilization were successful. However, only ∼10% of embryos were viable. Overall, our study demonstrates that the development of ARTs in individuals of Atelopus sp. (spumarius complex) bred in laboratory can be successful, which result in viable offspring through in vitro fertilization. Our study provides a baseline for assisted breeding protocols applicable to other harlequin frogs of the genus Atelopus.

Conservation of rare wild‐living cattle Bos taurus (L.): coat colour gene illuminates breed history, and associated reproductive anomalies have not reduced herd fertility

AbstractCoat colour patterns are important characteristics of many cattle breeds and in some are determined by a chromosomal translocation, which can have the pleiotropic effect of testicular hypoplasia. We test the hypothesis that this variant, known as the Cs29 allele, is prevalent in the ancient Chillingham White Cattle (Bos taurus) of north‐east England. Its distribution may provide insights into breed history, and as it is associated with reproductive anomalies, we investigated the issues of subfertility in this cattle herd which are of clear relevance to its conservation. We report that the cattle are homozygous for the translocation. As it is also known in other breeds of the England‐Scotland border and northern Irish area, namely the White Galloway and Irish Moiled, and in the Northern Finncattle and Swedish Mountain, we further hypothesize a Scandinavian connection for White Galloway, Irish Moiled and Chillingham cattle. We present unpublished data showing testicular hypoplasia to be present at Chillingham. Sperm quality is also known to be very low, and the question arises as to how the highly inbred (Fis = 0.92) Chillingham cattle have continued to survive in spite of these reproductive anomalies. Surprisingly, herd fertility has not declined over the last 160 years, and in the light of behavioural data, we propose this is probably because of multiple mating, there being no castration and only welfare culling in this herd. These findings have wide relevance, particularly for breeds of conservation importance and probably for other bovine species, because male subfertility, frequent in cattle generally, could restrict the choice of bulls for representation in gene banks with consequent risk of loss of lineages. Research on heterospermic insemination or multiple mating of cattle could, therefore, be a useful complement to the development of assisted reproduction technologies for cryoconservation.

CircSTK40 contributes to recurrent implantation failure via modulating the HSP90/AKT/FOXO1 axis

Increasing evidence has revealed a close relationship between non-coding RNAs and recurrent implantation failure (RIF). However, the role of circular RNAs (circRNAs) in RIF pathogenesis remains largely unknown. Microarray analyses were used to identify the differentially expressed circRNA-circSTK40. Functional experiments, including decidualization induction and terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) assay, were performed to determine the effects of circSTK40 on human endometrial stromal cells (ESCs). The interactions between circSTK40 and proteins were investigated by RNA pull-down, RNA immunoprecipitation, and co-immunoprecipitation (coIP) assays. We observed that circSTK40 expression was upregulated in the RIF midluteal-phase endometrial samples. circSTK40 overexpression in ESCs inhibited the decidualization process but concurrently enhanced cell survival during stress. Mechanistically, circSTK40 directly bound to HSP90 and CLU, thus functioning as a scaffold to block their interactions and hinder the proteasomal degradation of HSP90. The resulting high levels of HSP90 led to the activation of the AKT pathway and downregulation of FOXO1 expression. Inhibitors of AKT (MK-2206) and HSP90 (17AAG) both abolished the effects of circSTK40 overexpression in ESCs and increased the decidualization levels in a dose-dependent manner. Our findings indicate a novel epigenetic mechanism for RIF pathogenesis involving circSTK40 activity and provide a foundation for targeted treatments in patients with low endometrial receptivity.

Prevalence of pathogenic copy number variants among children conceived by donor oocyte

Development of assisted reproductive technologies to address infertility has favored the birth of many children in the last years. The majority of children born with these treatments are healthy, but some concerns remain on the safety of these medical procedures. We have retrospectively analyzed both the fertilization method and the microarray results in all those children born between 2010 and 2019 with multiple congenital anomalies, developmental delay and/or autistic spectrum disorder (n = 486) referred for array study in our center. This analysis showed a significant excess of pathogenic copy number variants among those patients conceived after in vitro fertilization with donor oocyte with respect to those patients conceived by natural fertilization (p = 0.0001). On the other hand, no significant excess of pathogenic copy number variants was observed among patients born by autologous oocyte in vitro fertilization. Further studies are necessary to confirm these results and in order to identify the factors that may contribute to an increased risk of genomic rearrangements, as well as consider the screening for genomic alterations after oocyte donation in prenatal diagnosis.

Canid Reproductive Biology: Norm and Unique Aspects in Strategies and Mechanisms.

Simple SummaryThe family Canidae, composed of dog-like species such as wolves, foxes, and jackals, demonstrates a significant variety in reproductive biology. In general, female canids experience very long periods of ovarian inactivity during the year; however, there are diverse patterns with regard to seasonality between species, as well as within an individual species depending on geographic region or housing status. Understanding of these differences is critical to the development of assisted reproductive technologies for canid conservation efforts. This review summarizes the current knowledge of canid reproduction, including reproductive cyclicity, seasonal breeding, male sperm traits, and recent developments in assisted reproductive technologies for canids.The reproductive physiology of canids is unique compared to other mammalian species. Specifically, the reproductive cycle of female canids is characterized by extended periods of proestrus and estrus followed by obligatory diestrus and protracted ovarian inactivity (anestrus). Although canid reproduction follows this general pattern, studies have shown variations in reproductive biology among species and geographic regions. Understanding of these differences is critical to the development of assisted reproductive technologies including estrus induction, gamete rescue, and embryo production techniques for canid conservation efforts. This review summarizes current knowledge of canid reproduction, including estrus cyclicity, seasonality, and seminal traits, with the emphasis on species diversity. The application of reproductive technologies in wild canid conservation will also be discussed.

The rise of non-traditional pregnancies through assisted reproductive technologies

This article explores how the development of assisted reproductive technologies (ARTs), arisen from in vitro fertilisation, have perpetuated an increase in non-traditional pregnancies ( Franklin, 1997 ). This article discusses what this increase means for midwifery practices and what care midwives may need to consider for such pregnancies. The discussions in this article are based on triangulated findings from a three-phase research design. The research consisted of an online mixed-methods survey of 521 citizens of the UK, semi-structured interviews with experts and professionals who speak to the future of ARTs, and structured interviews with people who are affected by genetic conditions. Findings reveal that applications of ARTs are anticipated to continue to expand with the potential introduction of genome editing technologies to reproductive choices for the prevention of disease. Thus, this article concludes that because of this expansion, the rise of non-traditional pregnancies is likely to continue, and that midwives may benefit from considering the implications the rise could have for midwifery care.

104 The effect of interspecific hybridization on the development of testes in genus Ovis males

Interspecific hybrids, especially males, are sterile or have reduced fertility. The structure and development of reproductive organs is species-specific. The study of development stages, morphology and biometric parameters in the male testes, the characterisation of species features, and the development of assisted reproductive technologies is of great importance. The aim of this research was to study morphometric parameters of the testes of interspecific hybrids of Argali with sheep of the Romanov breed compared with the original Romanov breed. The objects of research were purebred males of the Romanov breed (n=8) and hybrid males with the following genotypes: 1/4 Argali×3/4 Romanov (Group I, n=10), 1/8 Argali×7/8 Romanov (Group II, n=12). The collection of testes from purebred and hybrid animals was carried out at castration (age=12 months). Testicular tissue was fixed in Bouin’s solution. The following metrics were evaluated: weight and morphometric parameters of the testis, diameter of the seminiferous tubules, and the number of spermatogenic cells in the seminiferous tubule. Statistical analysis (comparison between groups with t-test and ANOVA) was performed using SPSS vs.15.0 (SPPS Inc.). The weight of testes in purebred Romanov males averaged 393±41g. In hybrid animals from groups I and II, this indicator was lower compared with that of purebred animals, by 58% and 41% (P&amp;lt;0.01), respectively. The average circumference of the testes in purebred animals and hybrids of groups I and II was 26±2, 19±3 and 20±3cm. The diameter of the seminiferous tubules in purebred animals and in hybrids of groups I and II was 215±3, 121±2 and 170±6µm, respectively. The number of spermatogenic cells inside the seminiferous tubules affects their size. In purebred animals, this value reached 90±3 cells. Hybrid animals in Groups I and II were inferior to their purebred peers in the number of spermatogenic cells in the seminiferous tubules by 31% (P&amp;lt;0.01) and 9%, respectively. The obtained results indicate the influence of individual characteristics of males on testicular morphology, including the number of spermatogenic cells in the seminiferous tubules. The studies were carried out with financial support from the Russian Science Foundation, grant No. 18-16-00079 and the Ministry of Science and Higher Education of the Russian Federation.

20 Development and Oct4/Cdx2 gene expression of Puma concolor, Leopardus geoffroyi, and Panthera onca hybrid embryos produced using domestic cat oocytes

The development of assisted reproductive technologies (ARTs) in Felidae species has been proposed to guarantee the genetic viability of species that are on the verge of extinction. Investigation of sperm cryopreservation and IVF have been studied in several Felidae species, but the lack of viable homologous oocytes makes assessment of sperm fertilization capacity after cryopreservation difficult. We propose to evaluate the capability of Puma concolor (Pc), Leopardus geoffroyi (Lg), and Panthera onca (Po) cryopreserved sperm to induce domestic cat oocyte fertilization and subsequent pre-implantation embryo development. Additionally, we evaluated blastocyst quality by analysing the gene expression patterns of pluripotency (Oct4) and differentiation (Cdx2) related markers using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). After ovariectomy, domestic cat (Felis catus, Fc) cumulus–oocyte complexes were matured invitro at 38.5°C in a humidified atmosphere of 6.5% CO2 and 21% O2 for 22h. Cryopreserved sperm from Pc, Lg, and Po were used for heterologous IVF. The sperm was frozen using Tes-Tris–based diluent (TEST), and survival, sperm motility, and acrosome integrity were evaluated. The quality post-thaw was satisfactory. The survival was high, sperm motility (65%) did not decrease during incubation post-thaw, and sperm with intact acrosome was 62% for all species. Briefly, straws were thawed by exposing them to air for 10s and then immersing in a 37°C water bath for 30s. The contents of the straws were poured into prewarmed sterile 1.5-mL microtubes. The sperm suspension was diluted (1:3 vol/vol) by the slow (drop by drop) addition of a modified Tyrode’s solution (Pope et al. 2006 Methods Molec. Biol. 254, 227-244). For IVF, invitro matured oocytes (n=635) were co-incubated with 0.5×105 mL−1 motile spermatozoa at 38.5°C in a humidified atmosphere of 6.5% CO2 and 21% O2 for 18 to 20h. Presumptive zygotes were cultured invitro in 50-µL drops of modified Tyrode’s medium in a humidified gas mixture of 5% CO2, 5% O2, and 90% N2 at 38.5°C. Cleavage was determined 48h post-fertilization and blastocyst stage was evaluated at Day 8 and stored in RNA-Later at −20°C until RT-qPCR analysis. Data were analysed by Fisher’s exact test using GraphPad Prism 6.0 (GraphPad Inc.), and differences were considered significant at P&amp;lt;0.05. We observed significantly lower cleavage rates of Po hybrid (hPo) embryos compared with hPc, hLg, and control Fc embryos (41% vs. 57%, 68%, and 58%, respectively). Additionally, we found that the blastocyst rate was higher using Lg sperm compared with sperm of other groups [hLg (50%) vs. hPc (29%), hPo (21%), and Fc (27%)].

Параметры спермограммы и генетические аномалии

The prevalence of genetic abnormalities in men with infertility is 5.8% (n = 9766), of which 4.2% are sex chromosome abnormalities and 1.5% are autosomal abnormalities. In the Russian Federation, this indicator varies from 4.72% (n = 539) (Novosibirsk) to 10.78% (n = 204) (St. Petersburg, the Research Institute of Obstetrics, Gynecology and Reproductology named after D.O. Ott) — the percentage of detectability probably depends on the concentration of patients in specialized institutions. Given the high frequency of genetic abnormalities in infertile men, it is necessary to correctly select diagnostic methods in accordance with the specifi c clinical situation. When the concentration of spermatozoa decreases to less than 10 million/ml, karyotyping is recommended; at a concentration of less than 5 million/ml, the search for AZF deletions is necessary; in severe disorders of spermatogenesis — the detection of mutations in the cystic fibrosis gene (CFTR). To confirm genetically determined asthenozoospermia, the electron microscopy of spermatozoa is required. Taking into account the development of assisted reproductive technologies, including the active use of intracytoplasmic sperm injection and the relationship of sperm pathology with severe genetically determined diseases, it is necessary to inform potential parents what risks to the health of future children the use of such material carries.