Possibility of elimination of pathogen from Babesia infected animals by means of embryo transfer

2014 ◽  
Vol 11 (2) ◽  
pp. 36-42
Author(s):  
P Erdenetogtokh ◽  
S Ganbat ◽  
Hiroshi Suzuki
Keyword(s):  
Embryo Transfer ◽  
Assisted Reproductive Technologies ◽  
Morphological Characteristics ◽  
Reproductive Technologies ◽  
Development Stage ◽  
Babesia Microti ◽  
Control Group ◽  
Mice Model ◽  
Economic Sectors

Babesia infections occur mainly in animals, and are transmitted by ticks. The severity of the diseases varies considerably depending on the species of Babesia involved as well as the immune response of the infected animal. In Mongolia infection produced by Babesia parasites is widely spread, provoking severe damage to the agricultural and economic sectors. Currently, strategies to control and prevent the infection are inefficient. Indeed, the necessity to look for suitable and accessible strategies to obtain animals free from the infection is needed. Currently, assisted reproductive technologies (ART) are used for the improvement of productivity in livestock. Moreover, embryo transfer seams to be useful approach to obtain clean embryos obtained from infected animals. Therefore, by using a mice model (ICR) infected with Babesia microti, an alternative method to obtain animals free from infection was examined. ICR mice at 8 weeks old were challenged with 0.2 ml of 1x107 IRBC/ml by i.p injection. After infection, superovulation was induced and then embryos were obtained and washed. Then, their development stage along with their morphological characteristics were monitored. In vitro embryos obtained from uninfected mice were used as a control group. The results indicate that the infection does not have any influence on pre-implantation embryonic development and morphological characteristics. Thus, we suggest that embryos obtained from infected animals might be useful for embryo transfer in order to improve productivity of livestock and reduce the risk of congenital infection. In summary, ART such as embryo transfer might be an useful technique in countries where Babesiosis is an endemic disease. DOI: http://dx.doi.org/10.5564/mjas.v11i2.214 Mongolian Journal of Agricultural Sciences Vol.11(2) 2013 pp.36-42

Mechanism of the adverse effect of hyaluronidase used for oocyte denudation on early development of bovine embryos

Zygote ◽  
2021 ◽  
pp. 1-5
Author(s):  
Shiori Ashibe ◽  
Kanade Irisawa ◽  
Ken Yokawa ◽  
Yoshikazu Nagao
Keyword(s):  
Treatment Group ◽  
Assisted Reproductive Technologies ◽  
Cumulus Cells ◽  
Reproductive Technologies ◽  
Control Group ◽  
Free Calcium ◽  
Parthenogenetic Development ◽  
Early Embryos ◽  
Bovine Oocytes

Summary Hyaluronidase is widely used in animal and human assisted reproductive technologies (ARTs) to remove cumulus cells around oocytes. However, adverse effects of hyaluronidase treatment, such as increased rates of degeneration and parthenogenesis, have been found after treatment of human and mouse oocytes. Currently, the mechanism(s) of the detrimental effects are unclear. The present study was initiated to identify the mechanism of adverse responses to hyaluronidase treatment in bovine oocytes and early embryos. Cumulus cells were removed from cumulus–oocyte complexes (COCs) with or without hyaluronidase and the oocytes were subjected to intracytoplasmic sperm injection (ICSI) or in vitro fertilization (IVF). Significantly lower rates of blastocyst formation were obtained in the hyaluronidase treatment group after ICSI (22.4%) and IVF (21.2%) compared with the non-hyaluronidase control groups: 36.1% after ICSI and 30.4% after IVF. Next, we examined the effect of hyaluronidase on parthenogenetic development rates and on the cytoplasmic levels of free calcium ions (Ca2+), reactive oxygen species (ROS) and reduced glutathione (GSH). No differences in parthenogenesis rates were found between treated and untreated groups. Ca2+ levels in oocytes from the hyaluronidase treatment group indicated using mean fluorescence intensity were significantly higher (68.8 ± 5.3) compared with in the control group (45.0 ± 2.5). No differences were found in the levels of ROS or GSH between the treated and untreated groups. We conclude that hyaluronidase might trigger an increase in Ca2+ levels in oocytes, resulting in a decreased potential for normal embryonic development.

Effectiveness and outcomes of embryo cryopreservation programs in assisted reproductive technologies

2014 ◽  
Vol 63 (4) ◽  
pp. 39-46 ◽  
Author(s):  
Yana Nikolayevna Kravchuk ◽  
Alla Stanislavovna Kalugina ◽  
Olga Vladimirovna Bystrova ◽  
Svetlana Aleksandrovna Shlykova
Keyword(s):  
Embryo Transfer ◽  
Assisted Reproductive Technologies ◽  
Negative Impact ◽  
Perinatal Outcomes ◽  
Reproductive Technologies ◽  
Embryo Cryopreservation ◽  
Control Group ◽  
Fresh Embryo Transfer ◽  
Art Programs ◽  
Thawed Embryo Transfer

Background. Embryo cryopreservation is an essential part of ART programs today. In recent years vitrification method is used increasingly widely. Purposes and tasks. To compare the effectiveness of ART programs using vitrified and fresh embryos, as well as different endometrial preparation regimes for frozen\thawed embryo transfer (modified natural cycle (MNC) and the preparatory hormone therapy(PHT)). To analyze the course of pregnancy and perinatal outcomes after vitrified embryo transfer. Materials and methods. We prospectively assessed the ART programs effectiveness and perinatal outcomes in 153 patients (I group), who underwent vitrified embryo transfer in 2011-2013 year. To prepare the endometrium for thawed embryo transfer in 83 patients PHT (Ia subgroup) and MNC in 70 patients (Ib subgroup) were used. Control group consisted of 70 patients, who underwent fresh embryo transfer. Results. The clinical pregnancy rate, birth rate and “take home baby” rate were not significantly different between the I (47,5 %; 30,9 %; 30,9 %) and II (53,0 %; 34,9 %; 32,5 %) groups, and between Ia (48,3 %; 28,4 %; 28,4 %) and IIb (46,6 %; 34,1 %; 34,1 %) subgroups. Complications during pregnancy and delivery, birthweight, length, Apgar score, congenital malformation rate did not differ significantly after vitrified and fresh embryo transfer. Conclusion. Vitrification is an effective method to achieve clinical results, comparable to native cycles. Application of PHT and MNC results in similar clinical outcomes. Transfer Vitrified embryo transfer does not have a negative impact on obstetric and perinatal outcomes when compared with native cycles.

scholarly journals Epigenetic Effects of Assisted Reproductive Technology in Human Offspring

2019 ◽  
Author(s):  
Wei Chen ◽  
Yong Peng ◽  
Xinyi Ma ◽  
Siming Kong ◽  
Shuangyan Tang ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Assisted Reproductive Technologies ◽  
Adult Life ◽  
Reproductive Technologies ◽  
Fresh Embryo Transfer ◽  
Vitro Fertilization ◽  
Nuclear Families ◽  
Genome Wide ◽  
Parental Background

AbstractThe births of more than 8 million infants have been enabled globally through assisted reproductive technologies (ARTs), including conventional in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) with either fresh embryo transfer (ET) or frozen embryo transfer (FET). However, the potential for elevated risks of ART-related disorders persists in adult life, and the underlying epigenetic mechanisms are largely uncharacterized. Here, we recruited 100 nuclear families and profiled the DNA methylomes, genome-wide histone modifications and transcriptomes to clarify the inherent extra risks attributable to specific ART procedures. We discovered that IVF-ET seemed to introduce less disturbance into the infant epigenome than IVF-FET or ICSI-ET did. Furthermore, we noted approximately half of the DNA methylomic changes in ART-conceived offspring could be explained by parental background biases. Through removal of the parental effect, we confirmed that ART per se would introduce minor DNA methylation changes locally. More importantly, we found that ART-induced epigenomic alterations were highly enriched in the processes which might contribute to increased incidence of preeclampsia during pregnancy and metabolic syndrome in offspring. Overall, our study provides an epigenetic basis for the potential long-term health risks in ART-conceived offspring that reinforces the need to review all methods of human ART.

scholarly journals Efficient marmoset genome engineering by autologous embryo transfer and CRISPR/Cas9 technology

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yukiko Abe ◽  
Harumi Nakao ◽  
Motoki Goto ◽  
Moe Tamano ◽  
Michinori Koebis ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Genome Engineering ◽  
Assisted Reproductive Technologies ◽  
Small Body ◽  
Common Marmoset ◽  
Reproductive Technologies ◽  
Reproductive Capacity ◽  
Primate Species ◽  
Novel Method

AbstractGenetic engineering of non-human primates, which are most closely related to humans, has been expected to generate ideal animal models for human genetic diseases. The common marmoset (Callithrix jacchus) is a non-human primate species adequate for the production of genetically modified animals because of their small body size and high reproductive capacity. Autologous embryo transfer (AET) is routinely utilized in assisted reproductive technologies for humans but not for experimental animals. This study has developed a novel method for efficiently producing mutant marmosets using AET and CRISPR/Cas9 systems. The embryos were recovered from oviducts of naturally mated females, injected with Cas9/guide RNA, and transferred into the oviducts of the donors. This AET method can reduce the time for in vitro culture of embryos to less than 30 min. This method uses an embryo donor as the recipient, thus reducing the number of animals and allowing for “Reduction” in the 3R principles of humane experimental technique. Furthermore, this method can utilize nulliparous females as well as parous females. We applied our novel method and generated the 6 marmosets carrying mutations in the fragile X mental retardation 1 (FMR1) gene using only 18 females including 14 nulliparous females.

97 PRELIMINARY DESCRIPTIVE STUDY OF EQUINE PLACENTA GENERATED AFTER TRANSFER OF IN VIVO- AND IN VITRO-PRODUCED EMBRYOS

2017 ◽  
Vol 29 (1) ◽  
pp. 156 ◽  
Author(s):  
A. Lanci ◽  
J. Mariella ◽  
B. Merlo ◽  
C. Castagnetti ◽  
E. Iacono
Keyword(s):  
Embryo Transfer ◽  
Intracytoplasmic Sperm Injection ◽  
Reproductive Technologies ◽  
Control Group ◽  
Placental Development ◽  
Group 3 ◽  
Group 2 ◽  
Group 1

Placental changes associated with artificial reproductive technologies have been described in several species, but little information is available in horses. Joy et al. (2012) reported that human placentas from intracytoplasmic sperm injection derived embryos were heavier and thicker than those produced after natural conception. Despite the most growing interest and efficiency of artificial reproductive technologies in equine species, only recently, Pozor et al. (2016) described placental abnormalities in pregnancies generated by somatic cell NT, but there are no studies on equine placenta generated by intracytoplasmic sperm injection and traditional embryo transfer. In the present preliminary study, macroscopic differences of placentas generated after transfer of in vitro- or in vivo-produced embryos were registered. Twelve Standardbred recipient mares with pregnancy generated after transfer of in vivo-derived (Group 1) and in vitro-derived (Group 2) embryos were enrolled; 10 Standardbred mares with pregnancy derived by traditional AI were included as control (Group 3). All pregnancies were physiological, and newborn foals were healthy. Mare age, parity, length of pregnancy, gross evaluation and weight of placenta, total length of umbilical cord (UC), length of UC, number of UC coils, foal sex, and weight at birth were registered. Collected data are listed in Table 1 and are expressed as mean ± standard deviation. Differences between groups were evaluated by 1-way ANOVA, and the difference in proportion of overweight placentas was evaluated with the Fisher test. The gross evaluation of placenta revealed 8/12 placentas (2/4 Group 1; 6/8 Group 2) were heavier than 11% (Madigan, 1997) due to oedema of the chorioallantois. No overweight placentas were registered in Group 3. In Group 1, 1/4 placentas had villous hypoplasia, and in Group 2, 1/8 placentas had cystic pouches on the UC. There were no significant differences among groups. However, the proportion of overweight placentas between Group 2 (6/8) and Group 3 (0/10) approached significance (P = 0.06). Although preliminary, the results of the present study suggest that production of equine embryos in vitro may lead to alterations in placental development. Several studies in cattle and sheep have suggested that alterations in the placentas of pregnancies derived from in vitro-produced embryos are related to effects of culture on epigenetic regulation. Less is known in the horse about the effects of in vitro embryo production on placental development; thus, further research in this area is necessary. Table 1. Characteristics of full-term placentas derived from AI or embryo transfer with in vivo- and in vitro-produced embryos

scholarly journals Long-Term Effects Following Fresh/Vitrified Embryo Transfer Are Transmitted by Paternal Germline in a Large Size Rabbit Cohort

Animals ◽  
2020 ◽  
Vol 10 (8) ◽  
pp. 1272 ◽  
Author(s):  
Ximo Garcia-Dominguez ◽  
José Salvador Vicente ◽  
María P. Viudes-de-Castro ◽  
Francisco Marco-Jiménez
Keyword(s):  
Embryo Transfer ◽  
Assisted Reproductive Technologies ◽  
Later Life ◽  
Reproductive Technologies ◽  
Early Embryo ◽  
Embryo Cryopreservation ◽  
Long Term Effects ◽  
Adult Body Weight ◽  
Female Germline

The concept of developmental programming suggests that the early life environment influences offspring phenotype in later life, whose effects may also be manifested in further generations. Valuable pieces of evidence come from the fields applying assisted reproductive technologies (ARTs), which deprive embryos of their optimal maternal environment and were thus associated with subsequent developmental deviations. Recently, we demonstrated that the in vitro manipulations during a vitrified embryo transfer procedure incurs a cumulative and transgenerational decline in the growth performance of the resulting offspring. Here, we provide a longitudinal study to investigate whether previous developmental deviations could be indistinctly paternally or maternally transmitted using crossbred mattings. Our findings revealed that early embryo manipulations through fresh and vitrified embryo transfer incurred paternally transmissible effects over the growth pattern and adult body weight, which seemed not inheritable via the female germline. Similar inheritable effects were observed after fresh and vitrified embryo transfer, suggesting that disturbing optimal embryo development through in vitro manipulations was the principal trigger of transmissible effects, rather than embryo cryopreservation per se.

176 IN VITRO MATURATION OF DOG OOCYTES IN CANINE FOLLICULAR FLUID

2014 ◽  
Vol 26 (1) ◽  
pp. 202
Author(s):  
K. Reynaud ◽  
S. Canguilhem ◽  
S. Thoumire ◽  
S. Chastant-Maillard
Keyword(s):  
Follicular Fluid ◽  
In Vitro Maturation ◽  
Assisted Reproductive Technologies ◽  
Cumulus Cells ◽  
Reproductive Technologies ◽  
Control Group ◽  
Limiting Factor ◽  
Cytoplasmic Maturation

In the canine species, assisted reproductive technologies, especially in vitro maturation (IVM) and IVF, are still ineffective. The main limiting factor remains the immaturity of the oocytes collected from anestrus ovaries. The ability of an oocyte to reach the MII stage in vitro is linked to the diameter of its follicle and anestrus oocytes, collected from small (<1 mm) follicles, are profoundly immature (De Lesegno et al. 2008). The objective of this study was to improve cytoplasmic quality by mimicking in vivo conditions; that is, to test the effect of pure preovulatory follicular fluid (FF) on survival and IVM rates of anestrus dog oocytes, in order to improve the nuclear and cytoplasmic maturation of these immature oocytes. Follicular fluids samples were collected from 54 Beagle bitches at 2 stages: before the LH peak (n = 23 bitches) and after the LH peak (n = 31 bitches). Only follicular fluid samples from large (>4 mm) follicles were collected and pooled by stage. Control oocytes were matured in 20% FCS/M199 medium. Groups of 5 oocytes were in vitro matured in 30 μL of follicular fluid, in half-area 96-well plates (5% CO2, 38°C). After 72 h of IVM, oocytes were denuded, fixed, and stained for DNA and tubulin before observation by confocal microscopy, and nuclear stages were classified as GV-A to GV-E, MI, and MII (Reynaud et al. 2012). A total of 460 oocytes were collected from 13 anestrus bitches and allocated to either the control medium (n = 155), the Pre-LH FF (n = 145) or the Post-LH FF (n = 160) groups. After 72 h of IVM, the morphology of the cumulus–oocyte complexes (COC) in the post-LH group was different from that of the others: cumulus cells appeared more compact and darker. Analysis of the nuclear stages showed that the degeneration rate was significantly higher (P < 0.05) in the post-LH group (58.7%) than in the pre-LH (40.9%) or in the control group (34.4%). No significant differences (P > 0.05) were observed between the 3 groups in the rate of immature GVA-B oocytes (36.4, 28.5, and 25.3% in the control, Pre-LH, and Post-LH groups, respectively), in the rate of meiotic resumption (GV-C/D/E, MI, MII stages, 44.4, 51.9, and 38.7% in the control, Pre-LH, and Post-LH groups, respectively). Metaphase II rates were not significantly different (12.1, 8.6, and 4.8% in the control, Pre-LH, and Post-LH groups, respectively). In conclusion, canine COC may survive when exposed to IVM in pure follicular fluid, but the degeneration rate was higher in the post-LH group. The presence of follicular fluid did not inhibit meiosis resumption, but did not significantly improve IVM rates. To better mimic in vivo conditions, IVM in a sequence of media, such as IVM in follicular fluid followed by IVM in oviducal fluid remains to be tested.

The combination of polymorphisms of genes PAI-1 -675 5G>4G, FXIII: Val34Leu G>T and ITGA2: 807 C>T in women with infertility can be considered as a predictor of failures of assisted reproductive technologies

2019 ◽  
Author(s):  
С.И. Сафиуллина ◽  
Я.Н. Котова ◽  
Е.С. Ворошилина ◽  
Н.А. Илизарова ◽  
Л.Ш. Ягудина ◽  
...  
Keyword(s):  
Assisted Reproductive Technologies ◽  
Treatment Effectiveness ◽  
Risk Groups ◽  
Reproductive Technologies ◽  
Fertility Treatment ◽  
Control Group ◽  
Vitro Fertilization ◽  
Ivf Failure ◽  
Pai 1

Введение. Результативность программ вспомогательных репродуктивных технологий остается неизменно низкой и не превышает 40 по числу положительных результатов хорионического гонадотропина человека и 23 по коэффициенту рождаемости. Актуальны новые способы увеличения эффективности лечения методом экстракорпорального оплодотворения (ЭКО) и вынашивания наступившей беременности. Цель исследования: на основании исследования генов полиморфизмов системы гемостаза выделить группы риска неудачных исходов программ ЭКО у женщин с бесплодием. Материалы и методы. Изучена когорта 130 женщин, планирующих лечение бесплодия методом ЭКО, и 49 женщин группы контроля. У всех женщин исследованы наиболее распространенные полиморфизмы системы гемостаза: FV: 1691 GA, FII: 20210 GA, FXIII: Val34Leu GT, FGB: 455 GA, ITGA2: 807 СT, ITGB3: 1565 TC, PAI 1: 675 5G4G методом полимеразной цепной реакции, выполнено сравнение их распространенности с аналогичными показателями контрольной группы. Проанализированы частоты встречаемости и значимости изученных полиморфизмов в 140 протоколах с переносом эмбрионов в зависимости от исхода. Результаты. У женщин с бесплодием, планирующих проведение программы ЭКО, не обнаружено достоверных различий в частоте распространенности изученных полиморфизмов системы гемостаза по сравнению с контрольной группой. Установлена достоверно высокая частота распространения триады полиморфизмов PAI1: 675 4G/4G, ITGA2: 807 СT и FХIII: Val34Leu GT у женщин с отрицательными исходами программы ЭКО по сравнению с положительными исходами. Заключение. Перспективно выделение группы риска неудач ЭКО на основании результатов генетического тестирования полиморфизмов системы гемостаза. Introduction. The effectiveness of assisted reproductive technology programs remains consistently low and does not exceed 40 in the number of positive results of human chorionic gonadotropin and 23 in terms of the birth rate. New ways of increasing the treatment effectiveness with in vitro fertilization (IVF) and carrying the new pregnancy are actual. Aim: to identify risk groups of IVF unsuccessful outcomes in women with infertility by studying of hemostasis genes polymorphisms. Materials and methods. We examined 130 women planning fertility treatment using IVF and 49 women as a control group. In all women we studied the most common hemostasis polymorphisms: FV: 1691 GA, FII: 20210 GA, FXIII: Val34Leu GT, FGB: 455 GA, ITGA2: 807 СT, ITGB3: 1565 TC, PAI1: 675 5G4G by polymerase chain reaction, and compared their prevalence with similar parameters of the control group. We analyzed the frequency of occurrence and significance of studied polymorphisms in 140 protocols with embryo transfer in dependence to outcome. Results. In women with infertility planning IVF program, there were no significant differences in the prevalence rate of studied hemostasis polymorphisms in comparison with the control group. We revealed significantly high frequency of 3 polymorphisms occurrence PAI1: 675 4G/4G, ITGA2: 807 CT and FХIII: Val34Leu GT in women with negative outcomes of IVF program in comparison with positive outcomes. Conclusion. Identification of risk groups of IVF failure based on the results of genetic testing of hemostasis polymorphisms is promising.

87 Assessment of pregnancy success following transfer of embryos produced in vitro using frozen - thawed semen from cloned and noncloned Bos indicus bulls

2019 ◽  
Vol 31 (1) ◽  
pp. 169
Author(s):  
O. Sebastián ◽  
F. Guerrero ◽  
R. Romero ◽  
F. Muñoz ◽  
A. Parlange ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Assisted Reproductive Technologies ◽  
Numerical Range ◽  
Bos Indicus ◽  
Reproductive Technologies ◽  
The State ◽  
In Vitro Production ◽  
Exact Test ◽  
Significant Difference

Assisted reproductive technologies (ART) continue to develop rapidly, allowing for the development of techniques to increase reproductive efficiency and contribute to the genetic improvement of cattle. Some of these techniques include in vitro production (IVP) of embryos and embryo transfer. These modern ART can help produce offspring with highly desirable characteristics. However, there is a lack of information on the percentage of pregnancies obtained following transfer (P/ET) of IVP embryos derived using semen of cloned Bos indicus bulls. The objective of this study was to compare embryo transfer results of IVP embryos created using frozen-thawed semen from 5 Brahman bulls (Bos indicus) with characteristics and genetics of high commercial value. The embryos were produced on two different dates, 45 days apart, using pooled oocytes harvested by ovum pickup from 15 Brahman cows at random stages of the oestrous cycle. Procedures for IVP were performed in a commercial laboratory (Genemex Internacional) in the state of Chiapas, Mexico. For IVF, conventional semen was used from 1 bull (B1) and his clone (B12), the grandson of B1 (B2), and from 2 nonrelated bulls (B3 and B4). A total of 100 embryos were transferred nonsurgically by a private practitioner on a ranch in the state of Campeche, Mexico. The recipients were commercial crossbred cows synchronized using a FTET program. On Day 0, recipients received an intravaginal device containing 1.9g of progesterone (CIDR) and 2mg of oestradiol benzoate IM. On Day 8, the CIDR was removed and cows received 25mg of dinoprost tromethamine, 200IU of eCG, and 0.5mg of oestradiol cypionate IM. Embryos were transferred on Day 17. The overall P/ET was 42% (42/100). The P/ET for IVP embryos produced with semen from bulls B1, B12, B2, B3, and B4 was 3/15 (20%), 3/8 (37%), 23/42 (55%), 8/20 (40%), and 5/15 (33%), respectively. The P/ET was numerically greater for embryos produced using semen from the cloned bull (37%; B12) compared to embryos produced using semen from the original noncloned bull (20%; B1), although this difference was not statistically significant (P=0.62, Fisher’s exact test). There was a significant difference (P&lt;0.05) for the P/ET obtained with embryos produced using semen from bulls B1 and B2, but results for the other bulls were not significantly different. As far as we know, this is the first scientific report in Mexico concerning the use and comparison of semen from cloned and noncloned bulls for the production and transfer of bovine IVP embryos. In general, a wide numerical range of P/ET using the different bulls was observed (i.e. 20-55%). In this preliminary study, there was no impact of using frozen-thawed semen from a cloned bull for IVP on P/ET. The results from this research can contribute to the study and development of ART to improve P/ET obtained using Zebu IVP embryos. However, further research with a larger numbers of animals is required to confirm whether using semen from cloned and noncloned Bos indicus bulls for IVP impacts pregnancy success following embryo transfer.

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