Cellular and molecular characterization of the impact of laboratory setup on bovine in vitro embryo production

2012 ◽  
Vol 77 (9) ◽  
pp. 1767-1778.e1 ◽  
Author(s):  
Dany Plourde ◽  
Christian Vigneault ◽  
Isabelle Laflamme ◽  
Patrick Blondin ◽  
Claude Robert
Keyword(s):  
Molecular Characterization ◽  
Embryo Production ◽  
Laboratory Setup ◽  
In Vitro Embryo Production ◽  
The Impact

123 In Vitro Embryo Production in Lyophilized In Vitro Culture Medium as a Method to Increase the Medium's Shelf-Life

2018 ◽  
Vol 30 (1) ◽  
pp. 201
Author(s):  
M. Rubessa ◽  
F. Salerno ◽  
D. Weisgerber ◽  
B. Gasparrini ◽  
B. Harley ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Shelf Life ◽  
Statistical Difference ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Freeze Dried ◽  
In Vitro Embryo Production ◽  
Α Level ◽  
The Impact

The worldwide production of livestock embryos requires stable medium with long shelf life. In this experiment, we evaluated the impact of the freeze-dried in vitro culture (IVC) medium (Mdry) on in vitro embryo production. We compared the standard IVC and Mdry media for cleavage rate and embryo production. Media solutions (10 mL) were aliquoted into 50-mL conical tubes and lyophilized to form a powder concentrate using a Genesis freeze-dryer (VirTis, Gardener, NY, USA). Lyophilization consisted of a constant cooling from 20°C to –10°C at a constant rate of 1°C/min with a 2-h hold at –10°C before sublimation at 0°C. Mdry medium were held at –80°C for 4 months. When the IVC medium was rehydrated, the pH were adjusted to 7.4. Abattoir-derived Holstein oocytes (n = 618, in 7 replicates) were in vitro matured and fertilized with sexed semen, according to standard procedures (Rubessa et al. 2011 Theriogenology 76, 1347-1355). Twenty hours after IVF, presumptive zygotes were cultured in SOF medium with 5% BS at 39°C with 5% CO2, 7% O2, and 88% N2. On Day 7, embryo yields were assessed. All recorded parameters were subjected to a Chi-Square Test 2 × 2. The parameters compared were percent cleavage, blastocysts, and embryos/cleaved. The α level was set at 0.05. All data were expressed as quadratic means and mean standard errors. The results (Table 1) showed not a statistical difference between control and Mdry. The Mdry had a higher percentage of cleaved zygotes (65.4% v. 53.4%) but not enough for a statistical difference. However, when we compared embryo production, there was no difference between treatments. The ratio between blastocysts and cleaved embryos was higher in the control group but not significant according to our selected α level. These results indicate that it is possible lyophilize IVC medium without interfering with the potential quality of the medium. Further studies will be needed to better understand the positive effect of the lyophilization on the cleavage rate. Table 1.Mean (SD in parentheses) percentage cleavage and blastocysts

scholarly journals Hazelnut (Corylus avellana) vicilin Cor a 11: molecular characterization of a glycoprotein and its allergenic activity

2004 ◽  
Vol 383 (2) ◽  
pp. 327-334 ◽  
Author(s):  
Iris LAUER ◽  
Kay FOETISCH ◽  
Daniel KOLARICH ◽  
Barbara K. BALLMER-WEBER ◽  
Amedeo CONTI ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Corylus Avellana ◽  
Cd Spectroscopy ◽  
Mediator Release ◽  
Food Allergies ◽  
Ige Reactivity ◽  
Allergenic Activity ◽  
The Impact

In Europe, hazelnuts (Corylus avellana) are a frequent cause of food allergies. Several important hazelnut allergens have been previously identified and characterized. Specific N-glycans are known to induce strong IgE responses of uncertain clinical relevance, but so far the allergenic potential of glycoproteins from hazelnut has not been investigated. The aim of the study was the molecular characterization of the glycosylated vicilin Cor a 11 from hazelnut and the analysis of its allergenic activity. Although MALDI–TOF (matrix-assisted laser-desorption ionization–time-of-flight) MS showed that one of two potential glycosylation sites of Cor a 11 was glycosylated, CD spectroscopy indicated that recombinant and natural Cor a 11 share similar secondary structures. Thus to analyse the impact of the glycan residues of Cor a 11 on IgE binding, the allergenic activity of natural glycosylated Cor a 11 and recombinant Cor a 11 was compared. In addition, the IgE sensitization pattern to recombinant Cor a 11, Cor a 1, Cor a 2 and Cor a 8 of 65 hazelnut allergic patients was determined in vitro. The prevalence of IgE reactivity to hazelnut vicilin Cor a 11 was below 50%. Basophil histamine-release assays were used to determine the allergenic activity of both natural and recombinant Cor a 11 in comparison with Cor a 1, a birch (Betula verrucosa) pollen-related major hazelnut allergen. Both forms of Cor a 11 induced mediator release from basophils to a similar extent, indicating that the hazelnut allergic patients had cross-linking IgE antibodies binding to the protein backbone and not to carbohydrate structures. In comparison to Cor a 1, a 10000-fold higher concentration of Cor a 11 was required to induce similar basophil mediator release. In conclusion, the hazelnut vicilin Cor a 11 is a minor allergen both in regard to prevalence and allergenic potency, whereas its glycan does not contribute to its allergenic activity.

scholarly journals Impact of ovarian fetal tissue xenotransplantation associated with in vitro embryo production in annual genetic gain estimates for cattle weight traits

2020 ◽  
Vol 26 (2) ◽  
pp. 15
Author(s):  
Giovanna Faria de Moraes ◽  
RAFAELLA KELLY SILVA ◽  
Luiza Rodrigues Alves Abreu ◽  
Filipe Luiz Jannuzzi Valente
Keyword(s):  
Genetic Gain ◽  
Ovarian Tissue ◽  
Fetal Tissue ◽  
Mcmc Methods ◽  
Embryo Production ◽  
Generation Interval ◽  
Average Value ◽  
In Vitro Embryo Production ◽  
The Impact

The present paper aims to estimate the impact of the reductionof the generation interval, based on the model of fetal ovarian tissuexenotransplantation associated with in vitro embryo production toestimate the annual genetic gain of weight traits in a bovine herd. Weighdata and genealogy of Brahman animals were used to estimate the annualdirect genetic gain for weight at birth, weight at 120 days, at 210 days,and at 550 days of age. The genetic parameters were obtained by animalmodel, via Bayesian inference, by Markov Chain Monte Carlo (MCMC)methods. For the estimates, different generation intervals were defined asthe average value between the supposed maternal generation intervalsand the average sire generation interval based on the analyzed data (9.6years). The shortest generation interval considered was 5.1 years, outlinedfrom the average of 0.66 year (corresponds to 8 fetal months, anassumption defined based on the use of fetal ovarian tissuexenotransplantation associated with in vitro embryo production) and 9.6years (sire generation interval). The remaining intervals considered were6.7, 8.7 and 10.2 years. The estimate of annual direct genetic gain for allthe evaluated traits was higher when considering the generation intervaloutlined based on the model of fetal ovarian tissue xenotransplantationassociated with in vitro embryo production. The estimates were lower asthe generation interval increased. The use of fetal ovarian tissuexenotransplantation associated with in vitro embryo production, may favorthe reduction of the generation interval and successively generate apositive impact on the annual direct genetic gain estimates.

scholarly journals Laparoscopic Ovum Pick-Up Followed by In Vitro Embryo Production and Transfer in Assisted Breeding Programs for Ruminants

Animals ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 216
Author(s):  
Hernan Baldassarre
Keyword(s):  
Pregnancy Rate ◽  
Commercial Application ◽  
Embryo Production ◽  
Breeding Programs ◽  
Sheep And Goats ◽  
Embryo Recovery ◽  
Marker Selection ◽  
Holstein Calves ◽  
In Vitro Embryo Production

The potential of laparoscopic ovum pick-up (LOPU) followed by in vitro embryo production (IVEP) as a tool for accelerated genetic programs in ruminants is reviewed in this article. In sheep and goats, the LOPU-IVEP platform offers the possibility of producing more offspring from elite females, as the procedure is minimally invasive and can be repeated more times and more frequently in the same animals compared with conventional surgical embryo recovery. On average, ~10 and ~14 viable oocytes are recovered by LOPU from sheep and goats, respectively, which results in 3–5 transferable embryos and >50% pregnancy rate after transfer. LOPU-IVEP has also been applied to prepubertal ruminants of 2–6 months of age, including bovine and buffalo calves. In dairy cattle, the technology has gained momentum in the past few years stemming from the development of genetic marker selection that has allowed predicting the production phenotype of dairy females from shortly after birth. In Holstein calves, we obtained an average of ~22 viable oocytes and ~20% transferable blastocyst rate, followed by >50% pregnancy rate after transfer, declaring the platform ready for commercial application. The present and future of this technology are discussed with a focus on improvements and research needed.

scholarly journals SureSelect targeted enrichment, a new cost effective method for the whole genome sequencing of Candidatus Liberibacter asiaticus

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Weili Cai ◽  
Schyler Nunziata ◽  
John Rascoe ◽  
Michael J. Stulberg
Keyword(s):  
Whole Genome Sequencing ◽  
Molecular Characterization ◽  
Genome Sequencing ◽  
Whole Genome Sequence ◽  
Whole Genome ◽  
Genome Coverage ◽  
Metagenomic Sample ◽  
Liberibacter Asiaticus

AbstractHuanglongbing (HLB) is a worldwide deadly citrus disease caused by the phloem-limited bacteria ‘Candidatus Liberibacter asiaticus’ (CLas) vectored by Asian citrus psyllids. In order to effectively manage this disease, it is crucial to understand the relationship among the bacterial isolates from different geographical locations. Whole genome sequencing approaches will provide more precise molecular characterization of the diversity among populations. Due to the lack of in vitro culture, obtaining the whole genome sequence of CLas is still a challenge, especially for medium to low titer samples. Hundreds of millions of sequencing reads are needed to get good coverage of CLas from an HLB positive citrus sample. In order to overcome this limitation, we present here a new method, Agilent SureSelect XT HS target enrichment, which can specifically enrich CLas from a metagenomic sample while greatly reducing cost and increasing whole genome coverage of the pathogen. In this study, the CLas genome was successfully sequenced with 99.3% genome coverage and over 72X sequencing coverage from low titer tissue samples (equivalent to 28.52 Cq using Li 16 S qPCR). More importantly, this method also effectively captures regions of diversity in the CLas genome, which provides precise molecular characterization of different strains.

Selection of stallions for in vitro embryo production by ICSI in a commercial program

2012 ◽  
Vol 32 (7) ◽  
pp. 409 ◽  
Author(s):  
C. Herrera ◽  
P. Dufourq ◽  
M. Freije ◽  
I. Morikawa ◽  
J.E. Centeno ◽  
...  
Keyword(s):  
Embryo Production ◽  
Commercial Program ◽  
In Vitro Embryo Production ◽  
Selection Of

In vitro embryo production after microinjection and ovarian dynamics following transvaginal follicular oocyte aspiration

1995 ◽  
Vol 43 (6) ◽  
pp. 1129-1139 ◽  
Author(s):  
J.R. Gibbons ◽  
R.L. Krisher ◽  
S.K. Carlin ◽  
R.E. Pearson ◽  
F.C. Gwazdauskas
Keyword(s):  
Embryo Production ◽  
In Vitro Embryo Production ◽  
Oocyte Aspiration ◽  
Ovarian Dynamics

Cleavage Rate of in vitro Matured Oocytes Fertilized with Conventional Semen in Buffaloes

2021 ◽  
Author(s):  
M.H. Pitroda ◽  
K.P. Khillare ◽  
M.B. Amle ◽  
M.D. Meshram ◽  
A.B. Mali ◽  
...  
Keyword(s):  
Fertilization Rate ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Quick Recovery ◽  
Slaughter House ◽  
Maturation Rate ◽  
Aspiration Technique ◽  
Current Scenario ◽  
In Vitro Embryo Production

Background: In vitro embryo production in buffaloes has gained much importance in this current scenario due to ever increasing population and high demand of milk and meat. Slaughter house derived bubaline ovaries are a cheap and abundant source of cumulus oocyte complexes.Methods: Oocytes from the buffalo ovarian follicles were recovered by aspiration technique as it facilitates quick recovery. Total 155 ovaries were used in the present study. Surface follicles were measured using vernier calliper and categorized into three groups viz. less than 3 mm, 3-5 mm and greater than 5 mm based on follicular diameter and oocytes were processed for IVM, IVF and IVC using conventional non sorted semen.Result: Overall percentage of small, medium and large follicles in the ovaries were recorded as 16.29 ± 0.94%, 8.14±0.60%, 5.35 ± 0.76%, respectively. Overall recovery rate of COCs was 38%. The percentage of these oocytes were 16.74% (A), 15.25% (B), 25.26% (C), 18.33% (D) and 29.87% (E) respectively. Maturation rate of oocytes were 81.96 ± 2.70%. Fertilization rate was 74.98 ± 3.87%, Cleavage rate % was 40.84±2.51% and Blastocyst percentage was 21.57±1.75% respectively. Application of in vitro embryo production technique using slaughter house ovaries can salvage the genetic potential of bubaline species.

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