scholarly journals Oocyte Source and Hormonal Stimulation forIn VitroFertilization Using Sexed Spermatozoa in Cattle

2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
Giorgio A. Presicce ◽  
Jie Xu ◽  
Guochun Gong ◽  
Juan F. Moreno ◽  
Sanjeev Chaubal ◽  
...  
Keyword(s):  
Hormone Treatment ◽  
Intramuscular Administration ◽  
Blastocyst Development ◽  
Embryo Production ◽  
Hormonal Stimulation ◽  
In Vitro Embryo Production ◽  
Donor Oocytes

The aim of this study was to investigate the efficiency of in vitro embryo production in cattle utilizing sexed sperm from two bulls and oocytes recovered by OPU. Twenty donor animals were employed in eight OPU replicates: the first four OPU trials were conducted on animals without hormone treatment, and the last four were run on the same animals, following FSH subcutaneous and intramuscular administration. A higher rate of blastocyst development was recorded in stimulated, as compared to nonstimulated animals, (25.2% versus 12.8%, ). Ocytes derived from slaughterhouse (SH) ovaries were also fertilized with sperm from the same bulls. Overall, non-sexed sperm used with oocytes derived from SH ovaries was significantly more efficient for blastocyst development than was sexed sperm with these same SH derived oocytes and sexed sperm with stimulated donor oocytes (39.8% versus 25.0% and 25.2%, ). In conclusion, the use of sexed sperm with OPU-derived oocytes resulted in a significantly higher blastocyst development when donors were hormonally stimulated; furthermore, the level of efficiency achieved was comparable to that attained when the same sexed sperm was tested on oocytes derived from SH ovaries.

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 < 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 < 0.04), blastocyst rate as a percentage of oocytes (16.7 ± 2.9 vs. 11.5 ± 2.1; P < 0.01), and blastocyst rates as a percentage of cleaved structures (24.1 ± 3.8 vs. 11.5 ± 2.1; P < 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.

135 Use and dose of porcine follicle-stimulating hormone for ovarian superstimulation prior to ovum pickup and in vitro embryo production in pregnant Holstein heifers

2019 ◽  
Vol 31 (1) ◽  
pp. 192
Author(s):  
R. V. Sala ◽  
L. C. Carrenho-Sala ◽  
M. Fosado ◽  
E. Peralta ◽  
D. C. Pereira ◽  
...  
Keyword(s):  
Limited Information ◽  
Dominant Follicle ◽  
Blastocyst Development ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Oocyte Competence ◽  
Treatment Groups ◽  
Oocyte Recovery ◽  
In Vitro Embryo Production

The benefit of superstimulation with exogenous FSH before ovum pickup for in vitro embryo production has been the subject of significant controversy. In addition, there is limited information on different dose regimens. Thus, the objective of the present study was to evaluate the effect of dose of porcine (p)-FSH during superstimulation before ovum pickup (OPU) on in vitro embryo production in pregnant heifers. Pregnant Holstein heifers (n=36) were assigned to a complete 3×3 crossover design. Three treatment groups were evaluated as follows: p-FSH 0mg (FSH0), p-FSH 160mg (FSH160) and p-FSH 300mg (FSH300). Three sessions of OPU were performed on each animal at 48, 62 and 76 days of gestation, with a washout interval between sessions of 14 days. Follicular wave emergence was synchronized by dominant follicle removal. Heifers in the FSH0 group received no further treatment, whereas the remaining groups received a total of 4 injections 12h apart as follows: FSH160 (48.0, 42.7, 37.3 and 32.0mg) or FSH300 (90.0, 80.0, 70.0 and 60.0mg), beginning 36h after dominant follicle removal. Ovum pickup was performed in all heifers 40h after the last p-FSH injection. Heifers were subjected to OPU for oocyte recovery, and number of follicles was determined. Recovered oocytes were processed and in vitro embryo production performed. Differences between treatment groups were evaluated by generalized linear mixed models. Data are presented (Table 1) as mean±standard error of the mean. There was no effect of days in gestation for any of the outcomes evaluated (P>0.05). Follicle numbers at the time of oocyte recovery were different (P<0.01) between groups. Heifers in the FSH300 group had a greater (P<0.05) number of medium, large and total follicles than heifers in the FSH0 group, whereas heifers in the FSH160 were intermediate. Total number of recovered, viable and cleaved oocytes were greater (P<0.01) in FSH300- than in FSH160- and FSH0-treated heifers. Cleavage rate and blastocyst development rate were not different (P>0.10) between groups. The number of grade 1 and 2 blastocysts was greater in FSH300- than in FSH160- and FSH0-treated heifers (P<0.03). In summary, the use of 300mg of p-FSH before OPU in pregnant heifers increases the number of follicles, oocytes and blastocysts produced per heifer with no detrimental effect on oocyte competence. Table 1.Ovum pickup and in vitro embryo production in pregnant heifers treated with different doses of porcine FSH

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 Exposure to follicular fluid during oocyte maturation and oviductal fluid during post-maturation does not improve in vitro embryo production in the horse

Zygote ◽  
2017 ◽  
Vol 25 (5) ◽  
pp. 612-630 ◽  
Author(s):  
Cécile Douet ◽  
Olivia Parodi ◽  
Nicola Antonio Martino ◽  
Giovanni Michele Lacalandra ◽  
Michele Nicassio ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Follicular Fluid ◽  
Culture Conditions ◽  
Blastocyst Development ◽  
Embryo Production ◽  
Development Rates ◽  
Maturation Medium ◽  
In Vitro Embryo Production ◽  
Oviductal Fluid

SummaryMost wild equids and many domestic horse breeds are at risk of extinction, so there is an urgent need for genome resource banking. Embryos cryopreservation allows the preservation of genetics from male and female and is the fastest method to restore a breed. In the equine, embryo production in vitro would allow the production of several embryos per cycle. Intracytoplasmic sperm injection (ICSI) is used to generate horse embryos, but it requires expensive equipment and expertise in micromanipulation, and blastocyst development rates remain low. No conventional in vitro fertilization (IVF) technique for equine embryo production is available. The development of culture conditions able to mimic the maturation of the oocyte in preovulatory follicular fluid (pFF) and the post-maturation in oviductal fluid (OF) may improve embryo production in vitro. Our aim was to analyse the effect of in vitro maturation in pFF and incubation in OF on in vitro maturation of equine oocytes, fertilization using conventional IVF or ICSI, and embryo development after culture in synthetic oviductal fluid (SOF) or DMEM-F12. Oocytes collected from slaughtered mares or by ovum pick up were matured in vitro in pFF or semi-synthetic maturation medium (MM). The in vitro maturation, fertilization and development rates were not statistically different between pFF and MM. After in vitro maturation, oocytes were incubated with or without OF. Post-maturation in OF did not significantly improve the fertilization and development rates. Thus, in our study, exposure to physiological fluids for oocyte maturation and post-maturation does not improve in vitro embryo production in the horse.

scholarly journals Evidence of haptoglobin in the porcine female genital tract during oestrous cycle and its effect on in vitro embryo production

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Francisco A. García-Vázquez ◽  
Carla Moros-Nicolás ◽  
Rebeca López-Úbeda ◽  
Ernesto Rodríguez-Tobón ◽  
Ascensión Guillén-Martínez ◽  
...  
Keyword(s):  
Western Blot ◽  
Female Genital Tract ◽  
Oestrous Cycle ◽  
Control Group ◽  
Blastocyst Development ◽  
Embryo Production ◽  
Or Efficiency ◽  
In Vitro Embryo Production ◽  
Proteomic Analyses

AbstractRecent evidence supports involvement of the acute phase protein haptoglobin in numerous events during mammalian reproduction. The present study represents an in-depth investigation of haptoglobin expression and secretion in the porcine oviduct and uterus, and assesses its effect on porcine in vitro embryo production. A systematic study was made of sows in different oestrous stages: late follicular, early luteal and late luteal stages. Relative haptoglobin mRNA abundance was quantified by RT-qPCR. In addition, expression of the protein was analysed by immunohistochemistry and the results were complemented by Western-blot and proteomic analyses of the oviductal and uterine fluids. In vitro porcine fertilization and embryo culture were carried out in the presence of haptoglobin. The results indicate that haptoglobin mRNA expression in the porcine oviduct and uterus is most abundant during the late luteal stage of the oestrous cycle. By means of Western blot and proteomic analyses haptoglobin presence was demonstrated in the oviduct epithelium and in the oviductal and uterine fluids in different stages of the oestrous cycle. The addition of haptoglobin during gamete co-incubation had no effect on sperm penetration, monospermy or efficiency rates; however, compared with the control group, blastocyst development was significantly improved when haptoglobin was present (haptoglobin: 64.50% vs. control: 37.83%; p < 0.05). In conclusion, the presence of haptoglobin in the oviduct and uterus of sows at different stages of the oestrous cycle suggests that it plays an important role in the reproduction process. The addition of haptoglobin during in vitro embryo production improved the blastocyst rates.

95 Successful Ovum Pick-Up-In Vitro Embryo Production in Lidia Cattle in France

2018 ◽  
Vol 30 (1) ◽  
pp. 187
Author(s):  
G. G. Lazo ◽  
S. Lacaze ◽  
D. Di Scala
Keyword(s):  
Transvaginal Ultrasound ◽  
Genetic Material ◽  
In Vitro Production ◽  
Blastocyst Development ◽  
Cleavage Rate ◽  
Embryo Production ◽  
In Vitro Embryo Production ◽  
Vitro Production ◽  
Maximum Humidity

Lidia cattle are a breed of Bos taurus that has been selected specially to produce bulls with the temperament and aggressiveness necessary to face a bullfighter in a ring. The genetic wealth of this fighting breed is divided into small lineages, traditionally called encastes, which has resulted in the risk of a loss of genetic variability (Ministerio de Medio Ambiente y Medio Rural y Marino, 2011; http://www.toroslidia.com/wp-content/uploads/2012/01/Programa-de-mejora-de-la-Raza-Bovina-de-Lidia.pdf). The technique to produce embryos in vitro may be a useful tool in the conservation of genetic material from this breed in a selection program. The aims of the study were to demonstrate the effectiveness of in vitro production of Lidia cattle embryos, and to evaluate variation in embryo production among males of the breed. Lidia cows, 7 to 13 years of age (n = 12), were used in an ovum pick-up (OPU)-in vitro production (IVP) program in the south of France. Ovarian superstimulation was induced with decreasing doses of pFSH (Stimufol; Reprobiol, Liège, Belgium) twice daily over 3 days (total dose: 350 µg). Transvaginal ultrasound-guided collection of cumulus–oocyte complexes (COC) was done 12 to 24 h after the last FSH injection. The COC were evaluated immediately after OPU and placed into 2.0-mL tubes (Corning Inc., Corning, NY, USA) containing 500 µL of maturation medium. A gas mix (5% CO2 in air) was injected into each tube and the tube was sealed tightly and placed in a portable incubator (Minitub, Tiefenbach, Germany) at 38.0°C for 12 h. On arrival in the Auriva IVP laboratory, tubes were opened and placed into an incubator with 5% CO2 at 38.5°C at maximum humidity to complete a 24-h maturation period. Semen was collected by electro-ejaculation previously from 5 different Lidia bulls (A, B, C, D, and E) and had been frozen by the same technique. The COC were fertilized with the frozen–thawed semen in TALP medium. Presumed zygotes were cultured in SOF medium (Minitub) to Day 7 (Day 0 = fertilization day) at 38.5°C in a 5% CO2, 5% O2, and 90% N2 atmosphere with maximum humidity. A total of 19 OPU/IVP sessions were performed, 5 cows were collected once, and 7 cows collected twice, and 143 COC were processed for in vitro embryo production. Blastocyst and expanded blastocyst numbers were recorded on Day 7. Oocyte recovery and embryo production by bull were analysed by ANOVA and blastocyst yield by Chi-square. The number (mean ± SEM) of oocytes allocated to each bull per IVP session was (P > 0.05): bull A (4.5 ± 1.9), bull B (5.8 ± 2.1), bull C (9.3 ± 2.5), bull D (6.5 ± 2.1), and bull E (7.0 ± 4.4). The cleavage rate differed among bulls (P < 0.05): bull A (4%), B (80%), C (89%), D (81%), and E (76%). The number (mean ± SEM) of blastocysts was lowest (P < 0.05) for bull A and highest (P < 0.05) for bull C (0, 3.7 ± 1.8, 7.0 ± 1.0, 4.3 ± 1.3, 4.7 ± 2.3 for bulls A to E, respectively). The blastocyst development rate (number of blastocysts/number of oocytes entering the IVF process) was also different among bulls (0, 63, 75, 65, and 67%, respectively; P < 0.05). Although there was a male effect on blastocyst production, our data demonstrate that successful in vitro embryo production in Lidia cattle is possible and suggests that this tool would be useful in a genetic program for the multiplication and the conservation of this breed.

159 Effect of Commercial Embryo Holding Medium on Development and Quality of Immature Bovine Oocytes

2018 ◽  
Vol 30 (1) ◽  
pp. 219
Author(s):  
M. Catteeuw ◽  
O. B. Pascottini ◽  
G. Opsomer ◽  
A. Van Soom
Keyword(s):  
Low Cost ◽  
Laboratory Work ◽  
Blastocyst Development ◽  
Embryo Production ◽  
Immature Oocytes ◽  
Developmental Capacity ◽  
In Vitro Embryo Production ◽  
Assess Quality ◽  
Bovine Oocytes

Bovine in vitro embryo production following ovum pick-up (OPU) in the field is hampered due to large time gaps between first and last OPU sessions. As oocytes will start immediate maturation, scheduling further manipulations makes the laboratory work laborious. There is a need for an easy and low-cost method that conserves the oocytes with full developmental capacity and allows scheduling laboratory work. In this regard, a commercial embryo holding medium (EHM; Syngro®; Bioniche Inc., WA, USA) was evaluated for conserving immature oocytes. Bovine immature oocytes (n = 2160) were collected from slaughterhouse animals by follicle aspiration and grouped per 60 and stored in 1 mL of EHM in 1-mL sterile glass osmometer tubes. Then, different temperatures [4°C, room temperature (RT), and 38.5°C] and different storage times (6, 10, 14 h) were assessed. After storage, oocytes were matured in TCM-199 supplemented with epidermal growth factor and gentamycin for 22 h. Fertilization was performed and zygotes (n = 1786) were cultured per 25 in 50 µL of SOF medium supplemented with 0.4% BSA and insulin, transferrin, and selenium. Control groups were included: immature oocytes (n = 1080) were not stored in EHM but immediately matured; fertilized and zygotes (n = 896) were cultured. Further, differential apoptotic staining was performed on a random subgroup of blastocysts to assess quality. Generalized fixed effect models were computed using R studio. Storage for 6 h showed a decrease in cleavage and blastocyst rate at 38.5°C (50 ± 3.9%; 11 ± 1.8%) compared with the control (78 ± 3.0%; 36 ± 2.8%). When increasing storage time, 38.5°C was not included; here, 4°C had a lower cleavage and blastocyst yield (47 ± 2.9%; 20 ± 3.3%) compared with the control (75 ± 2.5%; 41 ± 4.6%). For both 6 and 10 h, storage at RT resulted in similar cleavage (76 ± 3.4%; 74 ± 2.6%) and blastocyst rates (35 ± 2.7%; 40 ± 4.5%) as the control (P > 0.05). However, increasing storage to 14 h at RT decreased cleavage (61 ± 2.8%) and blastocyst yield (26 ± 2.5%) compared with the control (78 ± 2.4%; 39 ± 2.8%; P < 0.05). Evaluating embryo quality in all groups, no significant differences were found for any holding time or temperature of the EHM. To simulate OPU settings, EHM was also tested in 38 small groups of 10 immature oocytes that were subsequently matured, fertilized, and cultured. Based on the previous results, EHM storage was performed for 6 and 10 h at RT. Blastocyst development was not different between RT (19.8 ± 3.5%; 18.8 ± 3.6%) and the control (20.6 ± 3.6%; 18.3 ± 3.4%; P ≥ 0.05). To conclude, a commercial EHM can be used to conserve immature bovine oocytes without losing developmental capacity. Storage is recommended for no longer than 10 h and at RT in EHM. It opens new perspectives for practitioners, because this method is simple and low-cost; moreover, the start of maturation and subsequent in vitro embryo production process can be scheduled to avoid evening or night work at the laboratory.

Beneficial effects of melatonin on in vitro embryo production from juvenile goat oocytes

2018 ◽  
Vol 30 (2) ◽  
pp. 253 ◽  
Author(s):  
Sandra Soto-Heras ◽  
Montserrat Roura ◽  
Maria G. Catalá ◽  
Irene Menéndez-Blanco ◽  
Dolors Izquierdo ◽  
...  
Keyword(s):  
Embryo Quality ◽  
Cell Number ◽  
Blastocyst Development ◽  
Embryo Production ◽  
Melatonin Concentration ◽  
Oocyte Competence ◽  
Beneficial Effects ◽  
In Vitro Embryo Production ◽  
Ovarian Follicular Fluid

Melatonin is a universal antioxidant that improves in vitro embryo production in several species. The aims of this study were to determine the melatonin concentration in the ovarian follicular fluid (FF) of juvenile goats and the effect of melatonin during in vitro maturation (IVM) on embryo development. The FF melatonin concentration was 0.57­–1.07 × 10−9 M, increasing with follicular diameter. Oocytes were matured, fertilised and cultured under conventional conditions. Blastocyst development, embryo quality and levels of reactive oxygen species (ROS) and reduced glutathione were assessed. In Experiment 1 different melatonin concentrations (10−3, 10−7, 10−9, 10−11 M) were added to the IVM medium, which contained cysteamine as antioxidant, and no differences were observed. In Experiment 2, melatonin (10−7 M) was tested in the presence or absence of cysteamine (experimental groups: melatonin, cysteamine, melatonin + cysteamine, non-antioxidant). The melatonin group presented a higher blastocyst rate than the non-antioxidant group (28.9 vs 11.7%; P < 0.01) and a higher total cell number than the cysteamine group (225.1 vs 129.0; P < 0.05). Oocytes from the melatonin and cysteamine groups had lower ROS levels than those from the non-antioxidant group. This study shows that melatonin is an interesting tool for improving oocyte competence in juvenile goats as it increases embryo production and quality.

scholarly journals Detection of Brucella abortus during in vitro bovine embryo production

2021 ◽  
pp. 105-112
Author(s):  
A Akter ◽  
GK Deb ◽  
MFH Miraz ◽  
MA Kabir ◽  
SMJ Hossain ◽  
...  
Keyword(s):  
Brucella Abortus ◽  
Commercial Application ◽  
Bovine Embryo ◽  
Blastocyst Development ◽  
Embryo Production ◽  
Specific Pcr ◽  
Contagious Diseases ◽  
Biotechnological Approach ◽  
In Vitro Embryo Production

The in vitro embryo production (IVP) technology has emerged as a potential biotechnological approach to multiply genetically high yielding dairy cows. Its commercial application is increasing in many developed and developing countries over the years. Bangladesh livestock Research Institute (BLRI) adopted in vitro embryo production protocol from bovine ovaries of slaughterhouse. However, the risks of transmission of contagious diseases like Brucella abortus with embryos are not evaluated so far. Considering these facts, the present experiments were conducted to evaluate the efficiency of in vitro embryo production protocol with slaughterhouse ovaries as well as risk of contamination of produced embryos with Brucella abortus. To identify sources of contamination of embryos with Brucella abortus (if any), the laboratory water, different media used in the IVP process, semen, and follicular fluids were evaluated for confirmation of the organisms. In addition, vaginal swabs were collected from 2 buffaloes aborted due to suspected Brucella abortus infection. Molecular test were used to detect Brucella abortus contamination. Brucella abortus specific PCR product was not detected on agarose gel electrophoresis. The efficiency of IVP measured by cleavage and blastocyst development rates were 75.5±2.7% and 16.6±3.9%, respectively. The present study inferred that the in vitro produce embryos are free from Brucella abortus infection. Bang. J. Livs. Res. Vol. 27 (1&2), 2020: P. 105-112

160 EFFECT OF BSA SOURCES AND TYPES ON IN VITRO-PRODUCED LIVESTOCK EMBRYO DEVELOPMENT

2006 ◽  
Vol 18 (2) ◽  
pp. 188
Author(s):  
J. R. Dobrinsky ◽  
A. M. Paprocki ◽  
V. L. Chrostowski ◽  
C. M. Penfield ◽  
K. R. Rozeboom ◽  
...  
Keyword(s):  
Reproductive Biology ◽  
Embryo Development ◽  
Bovine Serum ◽  
Culture Medium ◽  
Production Systems ◽  
Culture Media ◽  
Blastocyst Development ◽  
Embryo Production ◽  
In Vitro Embryo Production

Bovine serum albumin (BSA) is a macromolecule supplement used in embryo and cell culture media. Other chemicals have been used as macromolecule substitutes in embryo culture with variable effectiveness. There are BSA products available that are defined in their disease status, collection method, and manufacturing process. They are compliant for use in raw form or in culture medium in the USA and EU. It was the purpose of this study to compare the effectiveness of highly defined and internationally compliant BSA with typically used BSA on in vitro-produced pig and cow embryo development. Pig oocyte-cumulus complexes were matured in two stages for 44 h in vitro. Semen from one boar of known high fertility was used across the study to fertilize mature oocytes. After 6-h co-incubation with sperm (50 motile sperm/oocyte), presumptive zygotes were cultured for 120 h in modified NCSU-23 containing 4 mg/mL of one of the following heat-shocked BSA fractions: Sigma A-7906 (control), Minitube Reproductive Biology Grade Fraction-V (RBG-V1, RBG-V2, RBG-V3), or Minitube Reproductive Biology Grade Fatty Acid-Free (RBG-FAF1). Across all treatments, Day 5 morulae were removed from BSA culture and placed into modified NCSU-23 with 10% fetal bovine serum (FBS) (no BSA) and cultured for 48 h. After 168-h total culture, the following blastocyst development was observed: Sigma A-7906, 12.3% (40/324); RBG-V1, 21.1% (36/171); RBG-V2, 19.0% (30/158); RBG-V3, 16.8% (27/161); and RBG-FAF1, 13.4% (21/157). These data show that culture medium supplemented with Minitube Reproductive Biology Grade BSA meets or exceeds (P < 0.05; ANOVA-GLM of SAS; SAS Institute, Inc., Cary, NC, USA) blastocyst development potential when compared to culture medium supplemented with undefined standard BSA preparations, such as Sigma A-7906. In vitro-matured cow cumulus-oocyte complexes were fertilized and cultured in one of two CR-1aa-based media. Oocytes were fertilized in IVF medium containing 6 mg/mL Sigma A-8806 (FAF) or 6 mg/mL Minitube RBG-FAF1. After 24 h in IVF medium, presumptive zygotes from a specific BSA-supplemented medium were cultured for 144 h in CR-1aa supplemented with their respective BSA (8 mg/mL). Day 6 morulae were removed from BSA culture and placed into CR-1aa with 10% FBS (no BSA) and cultured for 48 h. After 192-h total culture, the following blastocyst development from oocytes matured was observed: Sigma A-8806, 21.4% (88/411); RBG-FAF1, 18.9% (70/370). These data show that culture medium supplemented with Minitube Reproductive Biology Grade BSA meets blastocyst development potential (P > 0.05) when compared to culture medium supplemented with undefined standard BSA preparations, such as Sigma A-8806. The inclusion of internationally compliant BSA meets or exceeds blastocyst development rates in comparison to standard BSA preparations in common in vitro embryo production systems for swine and cattle. Although manufacturing differences remain the prominent variant in BSA sources and types, continued monitoring and documentation of BSA preparations tested in livestock in vitro embryo production systems will ensure a safe global supply of BSA products for future culture media production.

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