307 EFFECT OF BULL EPIDIDYMIS STORAGE CONDITIONS ON SPERM RESISTANCE AGAINST LIPID PEROXIDATION AND SUBSEQUENT IN VITRO EMBRYO PRODUCTION

2006 ◽  
Vol 18 (2) ◽  
pp. 261 ◽  
Author(s):  
M. Nichi ◽  
J. B. P. De Clercq ◽  
I. G. F. Goovaerts ◽  
V. H. Barnabe ◽  
P. E. J. Bols
Keyword(s):  
Lipid Peroxidation ◽  
Sperm Quality ◽  
Sperm Concentration ◽  
Membrane Lipid ◽  
Wilcoxon Test ◽  
Effect Of Temperature ◽  
Embryo Production ◽  
Sperm Membrane ◽  
In Vitro Embryo Production

Sperm recovery from the cauda epididymis can be a usefull tool in case of unexpected death of genetic high-value animals or endangered species or when the collection of sperm by other means becomes impossible. Studies indicate that the lower the temperature of epididymis storage, the better the sperm quality after collection (Kaabi et al. 2003 Theriogeneology 60, 1249-1259). One of the main factors that can negatively affect sperm viability during storage is lipid peroxidation, where sperm membrane resistance against reactive oxygen species (ROS) attacks is an important factor. The objective of this experiment was to study whether the temperature of epididymis storage following slaughter would have an influence on the membrane's resistance against lipid peroxidation and on the sperm cell's fertilizing capacity. Sixteen epididymides (from eight bulls) were collected after slaughter and divided into two groups, one stored at 4�C and the other at 37�C for 2 h, after which semen was collected from the caudae epididymides. Sperm concentration was measured and an aliquot containing 108 sperm cells was submitted to induced lipid peroxidation with ferrous sulfate and ascorbate (37�C; 2 h). Subsequently, thiobarbituric acid reactive substances (TBARS), as an index of lipid peroxidation, were measured according to a method previously described (Beorlegui et al. 1997 Andrologia 29, 37-42). A second aliquot of the sample was used for fertilization in a routine IVF-IVC set up in duplicate (24-h maturation, SOF culture medium in 5% CO2, 5% O2, and 90% N2). In vitro embryo production and level of TBARS were statistically analyzed using SAS (SAS Institute, Inc., Cary, NC, USA). TBARS levels were transformed to logarithm form in order to obey the residue normality being analyzed using PROC GLM. The percentage of blastocysts was analyzed using the Wilcoxon test. When compared to the samples stored at 4�C, semen of caudae epididymides stored at 37�C showed higher levels of TBARS and lower mean blastocyst rates (324.7 � 59.6 and 36.6 � 1.6 vs. 466.9 � 67.9 ng of TBARS/108 spermatozoa and 28.8 � 2.9%, respectively; P < 0.05). A negative correlation was found between TBARS and blastocyst rates (R = -0.43). The lower quality of sperm collected from epididymides maintained at higher temperatures may be related to a decrease in sperm resistance against lipid peroxidation which would further impair sperm fertilizing capacity. However, further studies are necessary in order to study the effect of temperature on the sperm membrane lipid profile, because the content of polyunsaturated fatty acids may be affected by temperature; this is an important factor relative to sperm membrane lipid peroxidation susceptibility (Ollero et al. 2000 Mol. Reprod. Dev. 55, 326-334). Another important factor is the epididymal environment because interactions between the sperm membrane and its surroundings can play an important role on the membrane's antioxidant protection.

Effect of storage for 24 h at 18°C on sperm quality and a comparison of two assays for sperm membrane lipid peroxidation

2010 ◽  
Vol 90 (3) ◽  
pp. 389-392 ◽  
Author(s):  
N. Am-in ◽  
R N Kirkwood ◽  
M. Techakumphu ◽  
W. Tantasuparuk
Keyword(s):  
Lipid Peroxidation ◽  
Membrane Permeability ◽  
Sperm Motility ◽  
Sperm Quality ◽  
Membrane Lipid ◽  
Membrane Lipid Peroxidation ◽  
Group A ◽  
Sperm Membrane ◽  
Fresh Semen

Boars having normal (71.1 ± 1.2%; n = 10) or low (35.12 &plusmn 3.9%; n = 10) sperm motility 24 h after collection were used, and semen was evaluated following storage in Beltsville Thawing Solution (BTS) for 24 h at 18°C. Sperm lipids were extracted and lipid peroxidation quantified. No differences were evident in fresh semen, but after 24 h, sperm motility, viability and membrane permeability in the low motility group were lower (P < 0.001) compared with the normal motility group. Sperm membrane lipid peroxidation was greater (P < 0.001) in the low motility group. A factor influencing sperm storability is membrane lipid peroxidation, which can be accurately assayed using a commercial kit.Key words: Boars, sperm motility, sperm quality, lipid peroxidation

145 Inhibitory effects of gossypol on bovine in vitro embryo production

2019 ◽  
Vol 31 (1) ◽  
pp. 197
Author(s):  
L. K. Hatamoto-Zervoudakis ◽  
M. F. Duarte Jr ◽  
T. F. Motheo ◽  
P. P. Tsuneda ◽  
J. T. Zervoudakis
Keyword(s):  
Embryo Development ◽  
Free Gossypol ◽  
Wilcoxon Test ◽  
Free Form ◽  
Bovine Embryo ◽  
Sodium Pyruvate ◽  
Embryo Production ◽  
Parametric Data ◽  
In Vitro Embryo Production

Cottonseed and its derivatives are frequently used in cattle feed as an effective dietary fibre supply and high protein and energy food source. However, the cotton plant contains gossypol, which in its free form induces male and female infertility. Therefore, this study aimed to evaluate the effect of gossypol supplementation on bovine in vitro embryo production. Ovaries were retrieved from slaughterhouses, and cumulus-oocyte complexes (COC) were recovered by follicular puncture. Based on free gossypol concentration present on the in vitro maturation, sperm capacitation, IVF and in vitro culture media, grades I, II and III COC (n=646) were divided in 3 treatments: 0μg mL−1 (control), 5μg mL−1 (G5) and 10μg mL−1 (G10). The COC were matured under a humidified atmosphere of 5% CO2 in air at 38.5°C for 24h in 90-μL droplets containing TCM-199 supplemented with 10% FCS, 0.2mM sodium pyruvate, LH, FSH, 75μg mL−1 amikacin, 17β-oestradiol. Each droplet corresponded to one replicate (n=14) and contained 15 to 18 COC. Matured COC and sperm were co-incubated in droplets (8-13 COC per 90μL) of TALP-IVF media supplemented with 6mg mL−1 BSA, 0.2mM sodium pyruvate, 30μg mL−1 heparin, 20 μM penicillamine, 10 μM hypotaurine, 1 μM epinephrine, 75μg mL−1 amikacin under a 5% CO2 humidified atmosphere at 38.5°C, for 20h. For IVF, non-sexed frozen-thawed semen was selected with Percoll® gradient. The resulting pellet was subjectively evaluated for motility and concentration and then diluted to final concentration of sperm mL−1 with fertilization medium. Presumptive zygotes were then cultured in 90-μL droplets of SOFaaci medium supplemented with 2.7mM myo-inosytol, 0.2mM pyruvate, 2.5% FCS (v/v), 5mg mL−1 BSA, 75μg mL−1 amikacin, and maintained for 8 days at 38.5°C in a humidified atmosphere with 5% CO2 in air. Cleavage, blastocysts production and hatching rates were evaluated at Days 3, 7 and 8, respectively. Data were submitted to ANOVA for parametric data and Wilcoxon test for non-parametric variables using the SAS software (SAS Institute Inc., Cary, NC, USA). Significance level was set at 5%. Cleavage rates of the control (81.05%) and G5 (71.85%) were higher compared with G10 (19.64%; P &lt; 0.0001). Blastocyst production was lower in G5 (12.18%) compared with control (30.35%), and the addition of 10μg mL−1 of free gossypol (G10) completely inhibited embryo development (0%; P &lt; 0.0001). As for the percentage of hatched blastocysts, the control (66.75%) had greater values compared with G5 (34.52%; P &lt; 0.0001). Thus, the addition of 5 and 10μg mL−1 of free gossypol are extremely hazardous for in vitro bovine embryo development. Whether these deleterious effects take place in a similar fashion during in vivo embryo production remains to be investigated.

scholarly journals Glycerophospholipids protect stallion spermatozoa from oxidative damage in vitro.

2021 ◽  
Author(s):  
Ashlee Jade Medica ◽  
Robert John Aitken ◽  
Garth Nicolson ◽  
Alecia Sheridan ◽  
Aleona Swegen ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Oxidative Damage ◽  
Unsaturated Fatty Acids ◽  
Sperm Quality ◽  
Membrane Lipid ◽  
Ros Generation ◽  
Sperm Viability ◽  
Commercial Preparation

Stallion sperm membranes comprise of a high proportion of poly-unsaturated fatty acids, making stallion spermatozoa especially vulnerable to peroxidative damage from reactive oxygen species generated as a by-product of cell metabolism. Membrane Lipid Replacement therapy with glycerophospholipid (GPL) mixtures has been shown to reduce oxidative damage in vitro and in vivo. The aims of this study were to test the effects of a commercial preparation of GPL, NTFactor® Lipids, on stallion spermatozoa under oxidative stress. When oxidative damage was induced by the addition of arachidonic acid to stallion spermatozoa, the subsequent addition of GPL reduced the percentage of 4-hydroxynonenal (4-HNE; a key end product of lipid peroxidation) positive cells (32.9±2.7 vs 20.9±2.3%; P≤0.05) and increased the concentration of 4-HNE within the spent media (0.026±0.003 vs 0.039±0.004 μg/mL; P≤0.001), suggesting that oxidized lipids had been replaced by exogenous GPL. Lipid replacement improved several motility parameters (total motility, 2.0±1.0 vs 68.8±2.9%; progressive motility, 0±0 vs 19.3±2.6%; straight line velocity, 9.5±2.1 vs 50.9±4.1 μm/s; curvilinear velocity, 40.8±10 vs 160.7±7.8 μm/s; average path velocity 13.4±2.9 vs 81.9±5.9 μm/s; P≤0.001), sperm viability (13.5±2.9 vs 80.2±1.6%; P≤0.001) and reduced mitochondrial ROS generation (98.2±0.6 vs 74.8±6.1%; P≤0.001). Supplementation with GPL during 17 oC in vitro sperm storage over 72 h improved sperm viability (66.4±2.6 vs 78.1±2.9%; P≤0.01) and total motility (53±5.6 vs 66.3±3.5%; P≤0.05). It is concluded that incubation of stallion spermatozoa with sub-mm-sized GPL micelles results in the incorporation of exogenous GPL into sperm membranes, diminishing lipid peroxidation and improving sperm quality in vitro.

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

scholarly journals 252 COMPARISON OF TWO SPERM SEPARATION PRODUCTS FOR USE IN BOVINE IVF

2005 ◽  
Vol 17 (2) ◽  
pp. 276 ◽  
Author(s):  
J. Pryor ◽  
S. Romo ◽  
D.D. Varner ◽  
K. Hinrichs ◽  
C.R. Looney
Keyword(s):  
Sperm Concentration ◽  
Embryo Production ◽  
Chi Square ◽  
Vitro Fertilization ◽  
Before And After ◽  
Chi Square Test ◽  
Group 2 ◽  
Live Sperm ◽  
Sperm Separation

In commercial bovine in vitro fertilization (IVF) companies, there is a continuous need to improve results. Efforts to maximize in vitro embryo production have included modifications in the use of sperm separation gradients. The development of commercially available sperm centrifugation gradients represents a new possibility of increasing the number of viable sperm that can be obtained from low concentration (fresh or frozen, sexed or unsexed) semen samples in order to improve the efficiency of the IVF system to make embryo production as efficient as possible. The objective of this study was to compare two different separation gradients, as follows: Group 1: Percoll (Sigma, St. Louis, MO, USA), in 45% and 90% gradients; Group 2: EquiPure (Nidacon, Gathenburg, Sweden), in top and bottom layers. Before and after separation, sperm were evaluated at 200× magnification for total motility, and then stained to assess viability at 400× with fast-green/eosin stain (Sigma). Sperm separation was performed using frozen/thawed semen from one bull. Semen was separated by centrifugation at 200g for 30 min in both density gradients. Results obtained from Groups 1 and 2 were compared by chi-square test. Sperm separation with Percoll yielded lower numbers of sperm (average sperm concentration after separation of 92 × 106, vs. 159 × 106 sperm/mL for EquiPure; P < 0.05) but resulted in higher motility (60% vs. 39%, respectively; P < 0.05) of separated sperm. Rates of live sperm cells were not significantly different between groups (69.5% vs. 70%, respectively; P > 0.1). These results indicate that the commercial separation medium EquiPure may be associated with higher sperm concentration levels but with lowered sperm motility when compared to Percoll for bovine sperm separation. However, Equipure provided similar percentages of live sperm when compared to Percoll, which is currently used in our laboratory.

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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