Frequency of exogenous hormone therapy impacts spermiation in male Fowler’s toad (Bufo fowleri)

2016 ◽  
Vol 28 (7) ◽  
pp. 995 ◽  
Author(s):  
Caitlin E. McDonough ◽  
Michelle W. Martin ◽  
Carrie K. Vance ◽  
Judith A. Cole ◽  
Andrew J. Kouba

Amphibians are experiencing a global extinction crisis and captive assurance colonies, along with reintroduction programs, are necessary to prevent further losses. Assisted reproductive technologies (ART), such as hormone-stimulated gamete collection and in vitro fertlisation (IVF), are conservation methods that can be used to increase reproductive output for breeding and reintroduction programs when animals fail to breed naturally. In order to maximise the production of offspring using ART, it is important to establish the physiological limitations on the frequency that hormone therapy can be used to collect gametes for IVF or assisted breeding. The present study examined the effects of the frequency of hormone-induced spermiation on sperm quantity and quality in Fowler’s toad (Bufo fowleri) by comparing four levels of hormone injection frequencies: twice a week, once a week, every other week, and every 3 weeks. Sperm release was induced with an intraperitoneal injection of 300 IU human chorionic gonadotropin (hCG). Spermatozoa were collected at three time points after injection (5, 7 and 9 h) and sperm concentration, motility and quality of forward progressive movement were measured. A significant decrease in sperm concentration (P < 0.01) was observed with the most frequent treatment (twice a week hormone injections). However, there was no negative effect of the treatments on sperm motility (P = 0.06) or forward movement (P = 0.06). We also observed a significant decrease in the concentration (P < 0.01), motility (P = 0.02) and quality of forward progressive movement (P = 0.01) of spermatozoa at the 9 h collection compared with earlier collection times. These results have clear implications for amphibian captive breeding programs, where more frequent hormone-induced spermiation could have a negative effect on male performance. We recommend that hormone injections be spaced a minimum of 2 weeks apart to optimise the health of the animals, assisted breeding, IVF or collection of gametes for genome resource banking.

2018 ◽  
Vol 156 (2) ◽  
pp. 95-105 ◽  
Author(s):  
Elif Diken ◽  
Matthias Linke ◽  
Jan Baumgart ◽  
Leonid Eshkind ◽  
Dennis Strand ◽  
...  

Although an essential component of assisted reproductive technologies, ovarian stimulation, or superovulation, may interfere with the epigenetic reprogramming machinery during early embryogenesis and gametogenesis. To investigate the possible impact of superovulation particularly on the methylation reprogramming process directly after fertilization, we performed immunofluorescence staining of pronuclear (PN) stage embryos with antibodies against 5mC and 5hmC. PN stage embryos obtained by superovulation displayed an increased incidence of abnormal methylation and hydroxymethylation patterns in both maternal and paternal pronuclear DNA. Subsequent single-cell RT-qPCR analyses of the Tet1, Tet2, and Tet3 genes revealed no significant expression differences between PN stage embryos from spontaneously and superovulated matings that could be causative for the abnormal methylation and hydroxymethylation patterns. To analyze the possible contribution of TET-independent replication-associated demethylation mechanisms, we then determined the 5mC and 5hmC levels of PN stage mouse embryos using immunofluorescence analyses after inhibition of DNA replication with aphidicolin. Inhibition of DNA replication had no effect on abnormal methylation and hydroxymethylation patterns that still persisted in the superovulated group. Interestingly, the onset of DNA replication, which was also analyzed in these experiments, was remarkably delayed in the superovulated group. Our findings imply an impact of superovulation on both replication-dependent and -independent or yet unknown demethylation mechanisms in PN stage mouse embryos. In addition, they reveal for the first time a negative effect of superovulation on the initiation of DNA replication in PN stage mouse embryos.


2016 ◽  
Vol 2016 ◽  
pp. 1-9 ◽  
Author(s):  
Salvatore Giovanni Vitale ◽  
Paola Rossetti ◽  
Francesco Corrado ◽  
Agnese Maria Chiara Rapisarda ◽  
Sandro La Vignera ◽  
...  

Assisted reproductive technologies (ART) have experienced growing interest from infertile patients seeking to become pregnant. The quality of oocytes plays a pivotal role in determining ART outcomes. Although many authors have studied how supplementation therapy may affect this important parameter for both in vivo and in vitro models, data are not yet robust enough to support firm conclusions. Regarding this last point, in this review our objective has been to evaluate the state of the art regarding supplementation with melatonin and myo-inositol in order to improve oocyte quality during ART. On the one hand, the antioxidant effect of melatonin is well known as being useful during ovulation and oocyte incubation, two occasions with a high level of oxidative stress. On the other hand, myo-inositol is important in cellular structure and in cellular signaling pathways. Our analysis suggests that the use of these two molecules may significantly improve the quality of oocytes and the quality of embryos: melatonin seems to raise the fertilization rate, and myo-inositol improves the pregnancy rate, although all published studies do not fully agree with these conclusions. However, previous studies have demonstrated that cotreatment improves these results compared with melatonin alone or myo-inositol alone. We recommend that further studies be performed in order to confirm these positive outcomes in routine ART treatment.


2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
O Somova ◽  
H Ivanova ◽  
N Sotnyk ◽  
K Kovalenko ◽  
I Feskova

Abstract Study question To evaluate the effect of platelet-rich plasma (PRP) testicular injections on spermogram parameters of men with severe oligoasthenoteratozoospermia (OAT). Summary answer The PRP testicular injections have beneficial effects on spermatogenesis and enhance sperm concentration and motility in infertile men with OAT. What is known already The use of PRP therapy in assisted reproductive technologies is debatable. Despite the recent evidence of its positive effects in promoting endometrial and follicular growth, data from clinical studies are limited. There are only a few papers on the effectiveness of PRP therapy in the treatment of male infertility and sexual dysfunction. In more detail, the influence of PRP on spermatogenesis was carried out only on experimental animals. Although the mechanisms of its action have not yet been clarified, it is assumed that PRP, containing many biologically active molecules, realizes its effect through the tissue regeneration and cell proliferation. Study design, size, duration This prospective study included 68 men (34.6±5.2) years old with severe OAT (≤4 million/ml, motility ≤30%, normal sperm morphology ≤1%) receiving hormonal and antioxidant (AO) therapy during 6 months before in vitro fertilization cycles. 33 of them were injected once with autologous PRP (0.5 ml in each testicle). Spermogram and testosterone level were analyzed before the treatment and in 3, 4 and 6 months after it. Participants/materials, setting, methods: Sperm concentration, motility and morphology in ejaculate of 33 men of PRP group were compared with those in the group of 35 men without PRP within 6 months of starting the treatment. Total and free testosterone level were measured in blood serum. PRP was prepared by centrifuging the patient’s own blood in the anticoagulant-containing tubes. The final concentration of platelets in the obtained sample was 950.000 – 1.250 000 cells in 1 ml. Main results and the role of chance 4 months after the PRP injection, sperm concentration and motility increased in 18 of 33 men of the PRP group compared with the baseline (before the treatment) – 4.2 (1.0; 6.9) vs 1.4 (0.1; 3.4) mln/ml (p &lt; 0.05) and 36.7 (30.6; 45.8) vs 17.7 (6.7; 28.2)% respectively (p &lt; 0.05).The maximum increase in sperm motility (but not in sperm concentration!) was observed in 24 men in 6 months – 49.6 (39.6; 56.4)% (p &lt; 0.05). Percent of morphologically normal spermatozoa in ejaculate slightly increased only in 12 men in that time period from 0–1% to 1–2%. The total testosterone level was 2.4 times higher than the baseline (31.6±7.2 vs 13.2±4.3 nmol/l, p &lt; 0.05), the free testosterone level was 1.8 times higher (14.5±3.5 vs 7.9±3.0 pgl/ml, p &lt; 0.05). Unlike the PRP group, in the group of men without PRP treatment, the sperm parameters did not changed compared with the baseline in 4 months after the starting hormonal and AO treatment. A significant increase of sperin concentration was observed only in 17 of 35 patients in 6 months. Sperm motility and percent of morphologically normal spermatozoa after the treatment did not differ from the baseline. Changes in the testosterone levels were similar to changes in PRP group. Limitations, reasons for caution Only young and middle-aged men were considered in the study. Large randomized controlled studies are required to confirm the PRP therapy efficacy and safety of f various fertility disorders. There are also no standardized protocols for PRP preparation. Wider implications of the findings: PRP therapy may have great potential for the treatment of male infertility and improving spermatogenesis. Optimization of methods of PRP preparation and dosage of testicular injections can enhance reproductive outcomes in assisted reproductive technologies. Trial registration number Not applicable


2012 ◽  
Vol 24 (1) ◽  
pp. 170 ◽  
Author(s):  
A. Kouba ◽  
E. Willis ◽  
C. Vance ◽  
S. Hasenstab ◽  
S. Reichling ◽  
...  

Species-specific differences in breeding strategies and physiology have limited the application of assisted reproductive technologies (ART) for critically endangered amphibians in captive assurance colonies. In 2006, the Memphis Zoo (MZ) initiated a program to develop ART for the critically endangered Mississippi gopher frog after natural breeding failed. Standard gamete collection and IVF developed by MZ for reproducing endangered toads such as the Wyoming or boreal toad were applied to the gopher frog with little success, especially hormonal therapy for sperm production. Using the leopard frog as a model species for Ranids, we tested the time and dose dependence of a luteinizing hormone releasing hormone analogue (LHRHa) and hCG on sperm quantity and quality. Initial findings from the leopard frog study were critical in designing the study on gopher frogs. Our objectives were to (1) compare 2 different hormones administered intraperitoneal (500 IU hCG vs 15 μg LHRHa) or their combination on spermiation in gopher frogs; (2) develop in vivo oocyte maturation and ovulation protocols using LHRHa (15 μg) and hCG (500 IU); and (3) transfer this technology to another institution as proof of principle. In gopher frogs, 100 and 83% of the males produced sperm in response to the LHRHa and the combination treatment, respectively, whereas only 16% responded to hCG alone. Sperm concentration peaked at 1 h post-administration for all treatments, with the LHRH/hCG cocktail treatment producing the highest concentration of sperm (mean = 4.6 × 106 ± 1.2 × 106 sperm mL–1, n = 6). No differences in motility were observed between treatments (P > 0.05). For females, a series of priming hormones of hCG and LHRHa were given several months before an ovulatory hormone regimen resulting in ovulation by 100% of the females (n = 6), whereas animals not primed failed to ovulate (n = 4). These 3 separate priming and IVF trials conducted between 2008 and 2010 resulted in each female laying ∼2000 eggs, with an average fertilization rate of 76% for inseminated eggs and hundreds of tadpoles produced. These IVF tadpoles represent the first captive reproduction of gopher frogs and highlight how ART can be applied to conservation and genetic management of threatened species. Subsequently, we tested our IVF protocols on gopher frogs at Omaha's Henry Doorly Zoo using fresh (collected on site) and chilled, shipped sperm from MZ. We collected 6169 eggs from 9 hormone-primed females with all animals ovulating. A portion of the total eggs ovulated were inseminated, resulting in 2401 fertilized eggs (38.9% of total eggs collected) across 18 different male–female pairings leading to viable tadpoles. In addition, sperm transferred overnight from the MZ produced 202/441 fertilized eggs (46%). The transfer of this technology and production of endangered amphibians using chilled, shipped sperm from live animals is a conservation milestone that can be applied to other captive breeding programs.


2020 ◽  
Vol 32 (2) ◽  
pp. 203
Author(s):  
N. Buzzell ◽  
S. Blash ◽  
K. Miner ◽  
M. Schofield ◽  
J. Pollock ◽  
...  

The objective of this study was to investigate a method of oviducal semen deposition as a strategy for producing offspring from poor-quality cryopreserved goat sperm. Invitro fertilisation (IVF) and AI are common assisted reproductive technologies used in small ruminants, but they have varied results in the goat. The use of poor-quality cryopreserved-thawed sperm (&lt;50% live/dead ratio at post-thaw) can decrease the rate of success. These procedures were performed in the month of November in Central Massachusetts in the United States (42° N). Seven 10-year-old dairy goats (Saanen, Toggenburg, and Alpine breeds) were synchronised and superovulated using a progesterone implant on Day 0, a prostaglandin injection at Day 7, two daily injections of 36mg of FSH ~12h apart on Days 12-15, and progesterone implant removal on Day 14 followed by an injection of 50µg of gonadotrophin-releasing hormone. Sperm deposition was performed on Day 17 (72 h after implant removal). The animals were anaesthetised using a standardised protocol, intubated, and maintained using isoflurane, and sterile prep was performed before a midline laparotomy procedure. Straws from a single ejaculate from a transgenic founder that was cryopreserved using a commercial two-step glycerol-egg yolk-based extender were used. A straw from this collection was post-thawed 30 days after collection and, using a commercial live/dead stain, 67% live sperm was determined. The optimal type of sperm prep and sperm concentration is unknown and may be dependent on sperm quality. Therefore, different gradient preps using Vitrolife SpermGrad at three volumes (1.5 (used on two animals), 1.0, and 0.5mL) as well as two volumes of IVF Bioscience Bovine BO-SemenPrep (4.0mL (used on two animals) and 2.0mL) were used. All five pellets were diluted in 1.0mL of IVF Bioscience Bovine BO-IVF media. Sperm concentrations ranging from 75×106 to 27×106 spermmL−1 were deposited into one oviduct; then, a 10:1 dilution was performed and 7.5×106 to 2.7×10 spermmL−1 were deposited into the contralateral oviduct. The depositions were performed just proximal to the uterotubal junction in a volume of 0.1mL of diluent via a tuberculin syringe attached to a 20-gauge needle. Two days following the procedure, oviducts were flushed postmortem from three of the seven randomly selected goats. All three had fertilised embryos, and nineteen 8-cell embryos were retrieved. Three of these embryos were surgically transferred to the distal uterine horn of a suitable recipient. The recipient became pregnant and produced a single offspring. The remaining four of seven goats were killed 41 days post-surgery. Two of the four goats were pregnant, with one carrying one fetus and the other carrying five fetuses. Further studies are needed to optimise this method, but these initial results indicate that oviducal semen deposition directly into the oviduct proximal to the uterotubal junction may be a suitable alternative for producing offspring from suboptimal cryopreserved-thawed goat sperm.


2019 ◽  
Vol 7 ◽  
pp. 2050313X1983415
Author(s):  
José Arturo Mora Rodríguez ◽  
Leonardo M Porchia ◽  
Felipe Camargo ◽  
Esther López-Bayghen

Male patients suffering from oligoasthenoteratozoospermia typically failed to achieve pregnancy, even with assisted reproductive technologies. Growth hormone and insulin-like growth factor 1 have been shown to regulate sperm quality parameters; therefore, the insulin-like growth factor 1 supplement could improve sperm parameters. Here, we determine the effect insulin-like growth factor 1 has on sperm parameters in a patient suffering from oligoasthenoteratozoospermia. A 47-year-old male was administered once a day 1.5 IU of insulin-like growth factor 1 by intradermal injection for 2 months. Seminogram analysis was performed before and after. Treatment with insulin-like growth factor 1 resulted in a 15.5-fold improvement in sperm concentration (1.1 × 106 vs 18.3 × 106 per mL), 71.4% change in volume (0.7 vs 1.2 mL), increased progressive motility (2% vs 43%), and the total volume of sperm with progressive motility (0% vs 23.6%). Here, we show that administering a daily dose of insulin-like growth factor 1 can improve sperm quality parameters.


Science ◽  
1907 ◽  
Vol 26 (664) ◽  
pp. 383-384
Author(s):  
H. A. Allard
Keyword(s):  

Author(s):  
Iu.V. Davydova ◽  
◽  
A.Y. Lymanskaya ◽  

The aim is to evaluate the effectiveness of the drug «Atoxil Plus» use in the group of pregnant women after assisted reproductive technologies (ART) with early mild toxicosis against the background of constipation. Materials and methods. The effectiveness of the drug «Atoxil Plus» (Orisil-Pharma) in 37 pregnant women with early mild toxicosis on the background of constipation was studied. The pregnant women in the study had no other comorbidities (pathology of the gastrointestinal tract, severe heart failure, infectious diseases, etc.). Taking into account that the definition of the quality of life indicator is an integral indicator of the effectiveness of treatment, the quality of life of pregnant women was assessed according to the «Gastrointestinal Simptom Rating Scale» (GSRS), a special questionnaire, before and after complex treatment of early toxicosis using the drug «Atoxil Plus». GSRS includes 15 questions to assess the following domains: the presence of abdominal pain, heartburn, constipation, diarrhea and signs of dyspepsia. The lower the results for the studied domains, the better. All pregnant women used the «Atoxil Plus» (2 g 3 times/day) for 10–14 days. Any general or local laxatives were excluded from the treatment regimen. Results. Treatment of early pregnancy toxicosis is important, given the loss of electrolytes, microelements, water and significant deterioration in the quality of life. High sorption capacity of Atoxil Plus in relation to microorganisms and bacterial toxins, the possibility of moderate therapeutic doses use due to the large active surface area, the increased binding rate of bacteria and their toxins provide quick therapeutic effect. Against the background of the «Atoxil Plus» enterosorbent use, not only does the constipation frequency not increase, which is very important during pregnancy, but the number of bowel movements is normalized due to the presence of lactulose in the medication, especially given the significant amount of gestagens pregnant women receive after ART. Conclusions. The non-invasive nature of the use of Atoxil Plus, its oral administration, high efficacy in pregnant women with early toxicosis, which manifests itself in improving the quality of life and general well-being, a high safety profile make it possible to recommend this drug as a treatment for early mild toxicosis against the background of constipation in pregnant women after ART. The study was conducted in accordance with the principles of the Declaration of Helsinki. The research protocol was approved by the Local Ethics Committee of the institution mentioned in the work. Informed consent of women was obtained for the research. The authors declare no conflict of interest. Key words: pregnant women, assisted reproductive technologies, constipation, treatment, sorbents.


2020 ◽  
Vol 7 (1) ◽  
pp. 10-18
Author(s):  
E. V. Kirakosyan ◽  
E. A. Sosnova

A review of 130 sources of world literature was conducted in the databases Scopus, Web of Science, MedLine, Cochrane CENTRAL, Cochrane Database of Systematic Reviews (CDSR), Database of Abstracts of Reviews of Effectiveness (DARE), EMBASE, Global Health, CyberLeninka, RSCI on the problem of the effect of combined oral contraceptives and hormone therapy during assisted reproductive technologies on the function of the hemostatic system in non-pregnant and pregnant women. In this review, we have summarized and analyzed information on the effect of sex hormones entering the womans body from the outside on individual links and the hemostatic system as a whole, determined the relationship between drug administration and thrombotic complications, optimized the sequence of diagnosis of conditions predisposing to thrombosis, substantiated the tactics of preparation and management patients taking exogenous hormones before and during pregnancy, including through in vitro fertilization.


1916 ◽  
Vol 50 (600) ◽  
pp. 757-763
Author(s):  
H. A. Allard

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