Relationship between the characteristics of frozen-thawed ram spermatozoa and in vitro embryo production

2001 ◽  
Vol 13 (3) ◽  
pp. 193 ◽  
Author(s):  
Lee H. A. Morris ◽  
W. H. Johnson ◽  
S. P. Leibo ◽  
B. C. Buckrell
Keyword(s):  
Logistic Regression ◽  
Sperm Motility ◽  
Membrane Integrity ◽  
Screening Tests ◽  
Breeding Programme ◽  
Embryo Production ◽  
Predictive Values ◽  
Commercial Breeding ◽  
In Vitro Embryo Production

To select rams suitable for ovine in vitro embryo production (IVP), the predictive values of the screening tests used to identify unsuitable rams need to be established. The present study examined some characteristics of frozen–thawed ram spermatozoa that might be evaluated routinely in a commercial breeding programme. These included sperm motility, plasma membrane integrity, morphology, and acrosome and capacitation status of the sperm population. Cryopreserved spermatozoa from four Dorset rams, which had previously satisfied the selection criteria for inclusion in a commercial breeding programme, were used for IVP. The overall contribution of the four rams and the ejaculates within each ram to the variability (R2) in the production of blastocysts was very small (2.1% and 2.5% respectively). The analysis of the sperm characteristics by logistic regression revealed a significant and positive association between total post-thaw sperm motility, viability and longevity with in vitro blastocyst production. However, there was no association between the other surface characteristics of the spermatozoa measured in this study with embryo production. Despite the absence of differences between the rams in the low incidence of polyspermic fertilization, the significant and detrimental effects of polyspermic fertilization on in vitro blastocyst production rates were quantified by logistic regression analysis. A large proportion of the variability within the IVP system was unaccounted for by the analysis of sperm and oocyte characteristics evaluated in this study. Thus, the identification of other factors contributing to the variability in the production of embryos in vitro warrants further investigation. No single sperm characteristic was sufficient to predict the ultimate outcome of blastocyst production. Rather, assessments of multiple characteristics within the IVP system are required to make accurate predictions.

365 QUALITY ASSESSMENT OF BOVINE CRYOPRESERVED SPERM AFTER SEXING BY FLOW CYTOMETRY AND ITS USE FOR IN VITRO EMBRYO PRODUCTION

2010 ◽  
Vol 22 (1) ◽  
pp. 339
Author(s):  
J. O. Carvalho ◽  
R. Sartori ◽  
G. M. Machado ◽  
G. B. Mourão ◽  
M. A. N. Dode
Keyword(s):  
Flow Cytometry ◽  
Sperm Motility ◽  
Membrane Integrity ◽  
Fertilization Rate ◽  
Computer Assisted ◽  
Blastocyst Development ◽  
Embryo Production ◽  
Acridine Orange Staining ◽  
In Vitro Embryo Production

Several studies using sex-sorted sperm by flow cytometry have shown that its fertility is reduced. Therefore, this study evaluated structural and functional characteristics of sperm sexed by flow cytometry. In addition, in vitro embryo production (IVP) and development was assessed when frozen-thawed unsorted and sex-sorted sperm from 4 Nellore bulls. Each ejaculate was separated into three fractions: non-sexed (NS), sexed for X-sperm (SX), and sexed for Y-sperm (SY). After thawing, each sample was analyzed for sperm motility by computer-assisted semen analysis (CASA, Berkeley, CA), sperm head agglutination, sperm morphology, membrane integrity by propidium iodide (PI) and 6-carboxy-fluorescein diacetate (CFDA) staining, acrosome integrity by peanut agglutinin (PNA), capacitation by chlortetracycline (CTC), and chromatin integrity by acridine orange staining. Then, the samples were placed in 45 : 90% (NS90) or 45 : 60% (NS60, SX, and SY) Percoll™ gradients. After Percoll™ centrifugation, sperm pellets were analyzed or used for IVP. All analyses were replicated independently three times. For IVP, 2,271 in vitro matured oocytes were used. To assess fertilization rate, presumptive zygotes were fixed and stained with lacmoid at 18 h post-insemination (hpi). Cleavage was evaluated at Day 2 (48 hpi) and blastocyst development at Days 6, 7, 8, and 9 of culture. Data were analyzed using generalized linear models. No differences (P > 0.05) were observed between SX and SY groups for e sperm variables evaluated either before or after Percoll™. However, non-sexed sperm had higher sperm motility, greater percentage of sperm with intact membranes, and greater percentage of live sperm with intact acrosomes than sexed sperm (P < 0.05). An effect of Percoll™ was observed in the non-sexed samples, with those submitted to 45 : 90% gradient having higher motility, greater percentage of cells with intact membrane, and lower recovery rate than those submitted to a 45 : 60% gradient. No differences among groups were observed for fertilization rate, being 74.0 ± 5.7, 63.2 ± 5.1, 67.2 ± 5.7, and 55.4 ± 5.9% for NS90, NS60, SX, and SY, respectively. Group NS90 showed a greater cleavage rate than did the SY group, while groups NS60 and SX had similar rates to the others. Blastocyst development rates on Day 6 to Day 9 were greater for group NS90. For example, on Day 8 the blastocyst rate was 34.9 ± 3.6, 22.2 ± 3.2, 18.1 ± 3.3, and 14.8 ± 2.9% forNS90, NS60, SX, and SY groups, respectively. All groups showed similar embryonic developmental stages on Day 6 to Day 9. Although sex-sorting affected sperm characteristics, it did not cause a decrease with in vitro fertility. However, differences in blastocyst rates between groups NS60 and NS90 indicated that the sperm selection protocol affected embryo production. Financial support: Embrapa Genetic Resources and Biotechnology.

scholarly journals Isolate® and Optiprep® minigradients as alternatives for sperm selection in bovine in vitro embryo production

2014 ◽  
Vol 94 (1) ◽  
pp. 35-42 ◽  
Author(s):  
L. L. Vianna ◽  
J. Pradieé ◽  
E. C. S. Santos ◽  
A. O. Gonçalves ◽  
L. F. M. Pfeifer ◽  
...  
Keyword(s):  
Sex Ratio ◽  
Sperm Motility ◽  
Semen Quality ◽  
Membrane Integrity ◽  
Sperm Selection ◽  
Selection Methods ◽  
Embryo Production ◽  
Embryo Survival ◽  
In Vitro Embryo Production

Vianna, L. L., Pradieé, J., Santos, E. C. S., Gonçalves, A. O., Pfeifer, L. F. M., Rheingantz, M. G. T., Dode, M. A. N., Vieira, A. D., Lima, V. F. H., Correa, M. N. and Pegoraro, L. M. C. 2014. Isolate® and Optiprep® minigradients as alternatives for sperm selection in bovine in vitro embryo production. Can. J. Anim. Sci. 94: 35–42. The objective of this study was to evaluate alternatives in small volumes to conventional gradient of Percoll® on semen quality, in vitro embryo production, sex ratio and embryo survival after vitrification. Thawed semen was randomly allocated to one of four density gradient selection methods: (1) conventional Percoll® (P), (2) MiniPercoll (MP), (3) MiniIsolate (MI), and (4) MiniOptiprep (MO). Sperm kinetics and quality were evaluated. Use of P, MP and MI gradients did not affect sperm motility (P>0.05). However, there was a decrease in total and progressive sperm motility in MO (70.8 and 51.3% vs. 87.3 and 69.5% for P; 87.3 and 73% for MP; 92.3 and 78.8% for MI; P<0.05). The MO had lower membrane integrity compared with P, MP and MI (39.7 vs. 70.5, 72.3, 63.8%, respectively, P<0.05). The percentage of blastocysts produced was higher in MI than in MP and MO (21.1 vs. 16.1 and 16.9%, P<0.05) and similar to P (18.4%; P>0.05). Sex ratio and embryo survival after vitrification were similar among groups (P>0.05). Semen selected by Isolate and Optiprep gradient, at the concentrations and small volumes used, demonstrated similar characteristics and in vitro embryo production to conventional Percoll® gradient.

Effects of Transport Medium Composition on in vitro Drug Permeation across Excised Pig Intestinal Tissue

2020 ◽  
Vol 10 ◽  
Author(s):  
Bianca Peterson ◽  
Henrico Heystek ◽  
Josias H. Hamman ◽  
Johan D. Steyn
Keyword(s):  
Medium Composition ◽  
Screening Tests ◽  
Intestinal Tissue ◽  
Predictive Values ◽  
Transport Medium ◽  
Intestinal Fluids ◽  
Lower Permeability ◽  
The Impact ◽  
Simulated Intestinal Fluids

Background:: Knowledge of the permeation characteristics of new chemical entities across biological membranes is essential to drug research and development. Transport medium composition may affect the absorption of compounds during in vitro drug transport testing. To preserve the predictive values of screening tests, the possible influence of transport media on the solubility of model drugs, and on the activities of tight junctions and efflux transporter proteins (e.g. P-glycoprotein) must be known. Objective:: The aim of this study was to compare the impact of different transport media on the bi-directional transport of standard compounds, selected from the four classes of the Biopharmaceutical Classification System (BCS), across excised pig intestinal tissue. Methods:: The Sweetana-Grass diffusion apparatus was used for the transport studies. Krebs-Ringer bicarbonate (KRB) buffer and simulated intestinal fluids in the fed (FeSSIF) and fasted (FaSSIF) states were used as the three transport media, while the chosen compounds were abacavir (BCS class 1), dapsone (BCS class 2), lamivudine (BCS class 3) and furosemide (BCS class 4). Results:: Abacavir exhibited lower permeability in both the simulated intestinal fluids than in the KRB buffer. Dapsone showed similar permeability in all media. Lamivudine exhibited lower permeability in FaSSIF than in the other two media. Furosemide exhibited improved transport with pronounced efflux in FaSSIF. Conclusion:: Different permeation behaviors were observed for the selected drugs in the respective media, which may have resulted from their different physico-chemical properties, as well as from the effects that dissimilar transport media components had on excised pig intestinal tissue.

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.

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.

PSX-A-10 Late-Breaking: Effects of paternal high energy diets on blastocyst development during in vitro embryo production in the bovine

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 363-363
Author(s):  
Dylan B Davis ◽  
Zachary Seekford ◽  
Mackenzie Dickson ◽  
Lucas Gonçalves ◽  
Samir Burato ◽  
...  
Keyword(s):  
High Gain ◽  
High Energy ◽  
Carcass Composition ◽  
Adaptation Period ◽  
Blastocyst Development ◽  
Embryo Production ◽  
Ad Libitum ◽  
In Vitro Embryo Production ◽  
To Receive

Abstract The objective of this study was to evaluate the effect of paternal high energy diets on blastocyst development during in vitro embryo production (IVP). Eight sires were stratified by body weight (initial BW = 946 ± 85 kg) and randomly assigned to the same diet (NEm = 2.10, NEg = 1.44, CP = 14.1%, NDF = 16.6%, DM basis) fed at two different inclusion rates while having ad libitum access to bermudagrass hay (NEm = 1.02, NEg = 0.45, CP = 10.2%, NDF = 71.6). After a 10-d adaptation period, sires were individually fed to receive 0.5% (MAINT) or 1.25% [High gain (HG)] of their BW daily for 67 days. At the end of the feeding period, semen was collected through electroejaculation and frozen. Antral follicles were aspirated from ovaries obtained from a slaughterhouse and utilized for IVP in 4 independent replicates (n = 2,227 total oocytes). Cleavage rates were evaluated 48 h after fertilization and blastocyst development rates were evaluated after 7 days of embryo culture. The proposed treatments successfully induced differences in BW gain (P &lt; 0.01; 2.28 vs -0.04 kg/d) and carcass composition (Rump fat: 1.63 vs. 0.41 cm, P = 0.08; Rib fat: 1.06 vs. 0.41 cm, P = 0.02; intramuscular fat: 3.5 vs. 3.0%, P = 0.36; for HG vs. MAINT sires, respectively). There was a significant decrease in cleavage rates (69.9 ± 2.5 vs. 65.0 ± 2.7; P &lt; 0.04), blastocyst rate as a percentage of oocytes (16.7 ± 2.9 vs. 11.5 ± 2.1; P &lt; 0.01), and blastocyst rates as a percentage of cleaved structures (24.1 ± 3.8 vs. 11.5 ± 2.1; P &lt; 0.01) for HG compared with MAINT sires. In conclusion, sires fed diets that induce highly anabolic conditions had impaired blastocyst development compared to sires fed a maintenance diet.

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