scholarly journals Low sperm to egg ratio required for successful in vitro fertilization in a pair-spawning teleost, Senegalese sole ( Solea senegalensis )

2021 ◽  
Vol 8 (3) ◽  
Author(s):  
Sandra Ramos-Júdez ◽  
Wendy Ángela González-López ◽  
Jhons Huayanay Ostos ◽  
Noemí Cota Mamani ◽  
Carlos Marrero Alemán ◽  
...  

Cultured Senegalese sole ( Solea senegalensis ) breeders fail to spawn fertilized eggs. The implantation of large-scale in vitro fertilization protocols, to solve this problem, has been frustrated by low production of poor quality sperm. Cultured females were induced to ovulate with a 5 µg kg −1 single injection of gonadotropin-releasing hormone agonist (GnRHa) and viable eggs (82.6 ± 9.2% fertilization) were stripped 41:57 ± 1:46 h after the injection. Sperm was collected from cultured males, diluted in modified Leibovitz and used fresh to fertilize the eggs. Males were not treated with hormones. A nonlinear regression, an exponential rise to a maximum ( R = 0.93, p < 0.0001) described the number of motile spermatozoa required to fertilize a viable egg and 1617 motile spermatozoa were sufficient to fertilize 99 ± 12% (±95% CI) of viable eggs. Similar, spermatozoa egg −1 ratios of 592 ± 611 motile spermatozoa egg −1 were used in large-scale in vitro fertilizations (190 512 ± 38 471 eggs). The sperm from a single male (145 ± 50 µl or 8.0 ± 6.8 × 10 8 spermatozoa) was used to fertilize the eggs. The mean hatching rate was 70 ± 14% to provide 131 540 ± 34 448 larvae per fertilization. The viability of unfertilized eggs stored at room temperature decreased gradually, and the sooner eggs were fertilized after stripping, the higher the viability of the eggs. The collection of sperm directly into a syringe containing modified Leibovitz significantly increased the percentage of motile spermatozoa (33.4 ± 12.2%) compared with other collection methods. The spz egg −1 ratios for Senegalese sole were at the lower end of ratios required for fish. Senegalese sole have a pair-spawning reproductive behaviour characterized by gamete fertilization in close proximity with no sperm competition. The provision of a large-scale in vitro fertilization protocol (200 µl of sperm per 100 ml of eggs) will enable the industry to operate sustainably and implement breeding programmes to improve production.

2020 ◽  
Author(s):  
Sandra Ramos-Júdez ◽  
Wendy Ángela González-López ◽  
Jhons Huayanay Ostos ◽  
Noemí Cota Mamani ◽  
Carlos Marrero Alemán ◽  
...  

AbstractCultured Senegalese sole (Solea senegalensis) breeders fail to spawn fertilised eggs and this bottleneck could be solved with the implementation of large-scale in vitro fertilisation protocols. However, low production of poor-quality sperm has frustrated the development of in vitro fertilisation protocols. Cultured females were induced to ovulate with a 5 µg kg-1 single injection of gonadotropin releasing hormone agonist (GnRHa) and good quality eggs (82.6 ± 9.2% fertilisation) were stripped 41:57 ± 1:46 h after the injection. Sperm was collected from cultured males, diluted in modified Leibovitz and used fresh to fertilise the eggs. A non-linear regression, an exponential rise to a maximum (R = 0.93, P < 0.0001) described the number of motile spermatozoa required to fertilise a viable egg and 1617 motile spermatozoa were sufficient to fertilise 99 ± 12% (± 95% CI) of viable eggs. Similar, spermatozoa egg-1 ratios of 592 ± 611 motile spermatozoa egg-1 were used in large-scale in vitro fertilisations with 190,512 ± 38,471 eggs. The sperm from a single male (145 ± 50 µL or 8.0 ± 6.8 × 108 spermatozoa) was used to fertilise the eggs. The mean hatching rate of the large-scale in vitro fertilisations was 70 ± 14 % to provide 131,540 ± 34,448 larvae per fertilisation. When unfertilised eggs were stored at room temperature the percentage of viable eggs decreased gradually and indicated the sooner eggs were fertilised after stripping the higher the viability of the eggs. The collection of sperm directly into a syringe containing modified Leibovitz significantly increased the percentage of motile spermatozoa (33.4 ± 12.2 %) compared to dilution in modified Leibovitz immediately after collection (6.6 ± 4.9 %). Senegalese sole have a pair-spawning reproductive behaviour characterised by external gamete fertilisation in close proximity with no sperm competition. The low spermatozoa egg-1 ratio required for maximum fertilisation was consistent with this reproductive behaviour and strategy. The provision of a large-scale in vitro fertilisation protocol (200 µL of sperm per 100 mL of eggs) will enable the industry to operate sustainably and implement breeding programs to improve production.


Complexity ◽  
2021 ◽  
Vol 2021 ◽  
pp. 1-15
Author(s):  
Yafei Li ◽  
Hongfeng Wang ◽  
Li Li ◽  
Yaping Fu

As one of the most effective medical technologies for the infertile patients, in vitro fertilization (IVF) has been more and more widely developed in recent years. However, prolonged waiting for IVF procedures has become a problem of great concern, since this technology is only mastered by the large general hospitals. To deal with the insufficiency of IVF service capacity, this paper studies an IVF queuing network in an integrated cloud healthcare system, where the two key medical services, that is, egg retrieval and transplantation, are assigned to accomplish in the general hospital, while the routine medical tests are assigned into the community hospital. Based on continuous-time Markov procedure, a dynamic large-scale server scheduling problem in this complicated service network is modeled with consideration of different arrival rates of multiple type of patients and different service capacities of multiple servers that can be defined as doctors of the general hospital. To solve this model, a reinforcement learning (RL) algorithm is proposed, where the reward functions are designed for four conflicting subcosts: setup cost, patient waiting cost, penalty cost for unsatisfied patient personal preferences, and medical cost of patient. The experimental results show that the optimal service rule of each server’s queue obtained by the RL method is significantly superior to the traditional service rule.


Author(s):  
Beth Atkinson ◽  
Emma Woodland

AbstractEmbryoGlue is available to patients at many in vitro fertilization clinics, usually at an additional cost. The efficacy of hyaluronan-enriched transfer medium (HETM) is supported by moderate quality evidence that indicates a significant improvement in clinical outcomes such as live birth rates for patients, including poorer prognosis women (i.e., maternal age factor [>35 years] and recurrent implantation failure). An increased multiple pregnancy rate has been reported with the use of HETM; therefore, a single embryo transfer policy should be considered in conjunction with the use of EmbryoGlue. There is no evidence to suggest that HETM has any detrimental impact, and therefore the use of HETM in clinics may be justified for a specific demographic of patients. Further robust evidence, in the form of meta-analyses or large-scale randomized controlled trials, is needed to build a sufficient consensus regarding the benefit of hyaluronan supplementation in embryo transfer media.


2018 ◽  
Vol 110 (4) ◽  
pp. 655-660 ◽  
Author(s):  
Samuel James Alexander Dobson ◽  
Maria Teresita Lao ◽  
Essam Michael ◽  
Alex C. Varghese ◽  
Kannamannadiar Jayaprakasan

2016 ◽  
Vol 28 (2) ◽  
pp. 245
Author(s):  
J. Stewart ◽  
M. Rubessa ◽  
K. Polkoff ◽  
S. Lotti ◽  
M. Wheeler

Bovine leukosis virus (BLV) is a pathogen that affects the bovine immune system and leads to lymphosarcoma, leukemia, decreased milk production, and increased culling rates in cattle. BLV-infected cattle herds can be found worldwide; in the United States, specifically, 38% of beef herds, 84% of all dairy herds, and 100% of large-scale dairy operation herds are infected (Buehring et al. 2014 Emerg. Infect. Dis. 5, 772–782). The main transmission between cattle in herds is affected leukocytes in blood. Several farm practices, such as dehorning, rectal palpation, and vaccinating can lead to the pathogen transmission. Due to international trade laws and biosecurity concerns, semen from a BLV-positive bull is illegal to sell within certain countries. Prior studies have looked at use of seropositive bulls in AI with little risk in affecting the dam (Burger et al. 2000 AVJR 60, 819). Other studies used semen that was artificially infected with the virus then used for IVF (Bielanski et al. 2000 Vet. Rec. 146, 255–256). The aim of this research was to evaluate naturally infected BLV donor semen using abattoir-derived oocytes and the possible contamination of in vitro-produced (IVP) embryos. Semen was collected and frozen by a private company. Three seropositive bulls and 1 negative control bull were selected. All positive bulls were selected based on availability of seropositive BLV status. Prior to the experiment, all bulls used were evaluated for motility, concentration, and morphology. The negative control was used in prior IVF experiments that produced acceptable results for use in this experiment. Frozen sperm were thawed at 37°C for 40 s and pelleted by centrifugation (25 min at 300 × g) on a Percoll discontinuous gradient (45–80% in Tyrode’s modified medium without glucose and BSA). The matured oocytes were purchased from DeSoto Biosciences (Seymour, TN, USA) and were IVF according to standard procedures (Rubessa et al. 2011 Theriogenology 76, 1347–1355). Using 200 oocytes per replicate, the 3 positive bulls and 1 control bull were allocated 50 oocytes per bull in each replicate. After 20 to 22 h of gametes co-incubation, zygotes were denuded and cultured for 7 days in SOF, followed by the evaluation of embryos (from tight morula until hatching blastocyst). Positive bull #1 produced and tested 48 embryos. Positive bull #2 produced and tested 41 embryos. Positive bull #3 produced and tested 46 embryos. The negative control produced and tested 55 embryos. Embryonic DNA extraction was performed using standard procedures (Sattar et al. 2011 Reprod. Domest. Anim. 46, 1090–1097). Nested PCR followed the Fechner evaluations methods (Fechner et al. 1996 J. Vet. Med. B 43, 621–630). To detect BLV presence, electrophoresis was used with a 2% agarose gel containing 0.1% ethidium bromide. A total of 190 embryos were evaluated that were produced in 3 replicates. All samples analysed showed no evidence of BLV. In conclusion, use of BLV seropositive donor semen showed no transmission of the virus upon IVF of the oocytes.


2008 ◽  
Vol 90 ◽  
pp. S398
Author(s):  
T. Okimura ◽  
M. Kuwayama ◽  
M. Kikuchi ◽  
T. Segawa ◽  
Y. Takehara ◽  
...  

Animals ◽  
2019 ◽  
Vol 10 (1) ◽  
pp. 3 ◽  
Author(s):  
Barbara Kij ◽  
Joanna Kochan ◽  
Agnieszka Nowak ◽  
Wojciech Niżański ◽  
Sylwia Prochowska ◽  
...  

Some human, bovine, and mouse in vitro fertilized (IVF) embryos with morphokinetic abnormalities such as fragmentation, direct cleavage, and cytoplasmic vacuoles have the potential to reach the blastocyst stage, which is related to a high potential for implantation. The latest techniques of embryo development observation to enable the evaluation and selection of embryos are based on time lapse monitoring (TLM). The aim of this study was to determine the frequency of morphological defects in feline embryos, their competence to reach the blastocyst stage, and their ability to hatch. Oocyte-cumulus complexes were isolated after the scarification of ovaries and matured in vitro. Matured oocytes were fertilized in vitro by capacitated spermatozoa. Randomly selected oocytes were observed by TLM for seven-to-eight days. Out of 76 developed embryos, 41 were morphologically normal, of which 15 reached the blastocyst stage. Of 35 abnormally developed embryos, 17 reached the blastocyst stage, of which six had single aberrations and 11 had multiple aberrations. The hatching rate (%) was 15.6% in normally cleaving embryos, 6.25% in embryos with single aberrations, and 3.33% in those with multiple aberrations. The present study reports the first results, found by using TLM, about the frequency of the morphological defects of feline embryos, their competence to reach the blastocyst stage, and their ability to hatch.


2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Jianhua Li ◽  
Jing Chen ◽  
Tiecheng Sun ◽  
Shuiwen Zhang ◽  
Tingting Jiao ◽  
...  

Abstract Background In vitro oocyte maturation (IVM) is being increasingly approached in assisted reproductive technology (ART). This study aimed to evaluate the quality of embryos generated by in-vitro matured immature follicles, as a guideline for further clinical decision-making. Methods A total of 52 couples with normal karyotypes underwent in vitro fertilization, and 162 embryos were donated for genetic screening. Embryos in IVF group were generated by mature follicles retrieved during gonadotrophin-stimulated in vitro fertilization (IVF) cycles. And embryos in IVM group were fertilized from IVM immature oocytes. Results The average age of the women was 30.50 ± 4.55 years (range 21–42 years) with 87 embryos from IVF group and 75 embryos from IVM group. The rate of aneuploid with 28 of the 87 (32.2%) embryos from IVF group and 21 of the 75 (28%) embryos from IVM group, with no significant difference. The frequency of aneuploid embryos was lowest in the youngest age and increased gradually with women’s age, whether in IVF group or IVM group and risen significantly over 35 years old. The embryos with morphological grade 1 have the lowest aneuploidy frequency (16.6%), and increase by the grade, especially in IVF group. In grade 3, embryos in IVM group were more likely to be euploid than IVF group (60% vs 40%, respectively). Conclusions IVM does not affect the quality of embryos and does not increase the aneuploidy rate of embryos. It is clinically recommended that women more than 35 years have a high aneuploidy rate and recommended to test by PGS (strongly recommended to screened by PGS for women more than 40 years). Women aged less than 35 years old for PGS according to their physical and economic conditions. Embryo with poor quality is also recommended to test by PGS, especially for grade III embryos.


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