196 COMPARING OOCYTE LIPID CONTENT WITH CIRCULATING CHOLESTEROL AND TRIGLYCERIDE LEVELS OF BOS TAURUS AND BOS INDICUS DONOR COWS

2008 ◽  
Vol 20 (1) ◽  
pp. 177 ◽  
Author(s):  
C. B. Ballard ◽  
C. R. Looney ◽  
B. R. Lindsey ◽  
J. H. Pryor ◽  
J. W. Lynn ◽  
...  

Intracellular lipid content has been found to be higher in Bos indicus oocytes compared with Bos taurus oocytes (Ballard et al. 2007 Reprod. Fertil. Dev. 19, 170). The objective of this study was to further determine whether there was a correlation between oocyte lipid content and circulating cholesterol (CHL) and triglyceride (TRG) levels of mature purebred Brahman (BR; n = 11) and English breed (EN; n = 13) beef cows. All donor cows were maintained on the same ration 30 days prior to and during the oocyte collection interval. Oocytes were collected from donors at 30-day intervals (3 replicates/female) by transvaginal ultrasound-guided aspirations. Blood samples were collected to evaluate total circulating CHL and TRG levels just prior to CIDR insertion and again prior to oocyte collection. Follicle-stimulating hormone (Folltropin-V; Bioniche, Belleville, Ontario, Canada) was given twice daily for 3 days in decreasing doses beginning on Day 4 after CIDR insertion, with oocyte recovery on Day 8. The mean number of follicles aspirated/donor and oocytes recovered/donor were 12 and 11 for the BR and 18 and 7 for the EN donors, respectively. Oocytes were matured in TCM-199 with 10% fetal bovine serum + bovine LH and bovine FSH (0.01 U mL–1). After 20 h, mature oocytes with a visible polar body (BR = 242/391 or 61%, and EN = 170/258 or 65%) were denuded by vortexing in Tyrodes HEPES medium + BSA (0.03 mg mL–1). Visual color scores (1 = light, 2 = medium, 3 = dark) were then assigned to the oocytes. Buoyancy of mature oocytes was then measured by using a sucrose step-density gradient column prepared with sucrose and Dulbecco's PBS. Results from the sucrose gradients ranged from 23% (high lipids) to 35% (lower lipids). The oocytes were stained with Nile Red and evaluated by fluorescent microscopy. Oocyte images were evaluated by using Scion Image software to calculate the Nile Red units (NRU; higher NRU = higher lipid content). Serum was analyzed (Texas Veterinary Medical Diagnostic Laboratory) for CHL and TRG concentrations. Data were analyzed by one-way ANOVA. There was no difference between visual color scores for oocytes from BR and EN donors. However, BR oocytes had significantly (P < 0.05) lower buoyant density and significantly higher mean NRU scores (32% and 173) compared with oocytes harvested from EN donors (33% and 152), respectively. Serum CHL and TRG levels were both significantly higher (P < 0.05) in the BR donors (150 and 33.2) compared with the EN donors (110 and 25.5), respectively. These results suggest that serum cholesterol and triglyceride levels may be predictive of donor oocyte lipid content.

2007 ◽  
Vol 19 (1) ◽  
pp. 170 ◽  
Author(s):  
C. B. Ballard ◽  
C. R. Looney ◽  
B. R. Lindsey ◽  
J. H. Pryor ◽  
J. W. Lynn ◽  
...  

Bos indicus embryos have a lower survival rate compared with Bos taurus after cryopreservation. It has been hypothesized that the lower survival rate is due to higher intracellular lipid content. The objective of this study was to determine if there is a difference in intracellular lipid content of oocytes from mature purebred Brahman and Angus cows. Donor females used in the study were maintained on pasture prior to the onset of the experiment and on a grain-supplemented hay ration during the study. Oocytes were collected from cows at 30-day intervals (3 aspirations/donor) by transvaginal ultrasound-guided oocyte aspirations (TUGA). Mature oocytes were evaluated using a sucrose step gradient procedure and Nile Red staining. FSH (Folltropin-V®; Bioniche Animal Health, Beltsville, Ontario, Canada) administration began on Day 4 of the estrous cycle (estrus = Day 0) twice daily for 3 days in decreasing doses (Brahman 232 mg and Angus 280 mg total), and on Day 8 oocytes were recovered. The mean number of follicles aspirated/donor and oocytes recovered/donor were 20 and 16.61 oocytes for the Brahman donors (n = 6) and 12 follicles and 7.06 oocytes/donor for the Angus donors (n = 10). Oocytes (individual donor basis) were then incubated in TCM-199 supplemented with 10% fetal bovine serum + bLH and bFSH (0.01 U mL−1) at 38.5°C. After 20 h, mature oocytes were denuded by vortexing for 3 min in HEPS + BSA (4 mg mL−1). Buoyancy was tested for individual mature oocytes using a sucrose step density gradient column prepared with sucrose and Dulbecco's PBS. Results from the sucrose gradients ranged from 23% sucrose (indicating high lipids) to 35% sucrose (indicating lower lipids). Oocytes recovered from the sucrose were fixed for 24 h in paraformaldehyde for evaluation with Nile Red stain. Oocytes were stained for 24 h, and then placed in Prolong® Gold (Invitrogen, Carlsbad, CA, USA) and evaluated under fluorescence. Oocytes images were evaluated using a Scion Image camera (Scion Corp., Frederick, MD, USA) to calculate mean (± SE) Nile Red units (NRU) (higher NRU = higher lipid content). Treatment groups were analyzed using one-way ANOVA. In summary, Brahman M-II oocytes had significantly lower (P ≤ 0.05) buoyant density, with a significantly higher mean NRU score, when compared with oocytes harvested from Angus donors (Table 1). Based on these results, Brahman oocytes have a higher intracellular lipid content then Angus oocytes. Table 1.Percent sucrose levels and Nile Red units for bovine oocytes from three replicates per donor


2013 ◽  
Vol 25 (1) ◽  
pp. 226 ◽  
Author(s):  
E. P. López-Damián ◽  
T. Fiordelisio ◽  
M. A. Lammoglia ◽  
M. Alarcón ◽  
M. Asprón ◽  
...  

Accurate evaluation of bovine embryos for assessing developmental stage and quality is critical to the success of any embryo transfer program. However, this evaluation process has been reported to be highly subjective in Bos indicus (BI) and can vary as much as 23% compared with that of Bos taurus (BT). These differences in assessment may be related to the quantity of lipid droplets (LD) within the embryo, which has been shown to have a negative effect in cryopreserving embryos. The aim of the present study was to characterize the number and size of LD in different developmental stages of fresh embryos from BI and BT and to compare LD across the three different embryo quality grades (1 = excellent or good, 2 = fair, and 3 = poor). Nonsurgical embryo collection was performed 7 days post-insemination in 10 BI and 10 BT females. Forty-eight embryos were evaluated for stage and grade using stereoscopic microscopy, processed for transmission electron microscopy, and stained with Nile red. Digitalized images were analyzed with ImageJ (National Institutes of Health, Bethesda, MD, USA), contour of lipid droplets were designed, and values of perimeter, area, and fluorescence intensity were assessed. Nonparametric statistical analysis (Mann–Whitney) was utilized. There was no difference in LD number for BT or BI for morulae and blastocyst; however, BI morulae presented larger LD compared with blastocyst stage embryos (286 µm2 v. 223 µm2; P < 0.05). Likewise, BI TF cells had more LD compared with inner cell mass (ICM) cells (48 v. 36; P < 0.05). BT TF cells exhibited larger LD compared with ICM cells (149 µm2 v. 128 µm2; P < 0.05), while BI embryos exhibited a larger area of LD in the ICM compared with the TF (591 µm2 v. 472 µm2; P < 0.05). In all embryos, BI contained more lipid droplets than BT (78 v. 49; P < 0.05). Across all quality grades (good, fair, and poor) there was no difference in the number of LD in BT embryos; however, BI grade-3 embryos presented more LD than grade-1 (36 v. 25). BT embryos LD were larger than BI LD (907 µm2 v. 625 µm2; P < 0.05). Fluorescence images showed higher arbitrary units of fluorescence (auf) for LD in BI. Compared with BT embryos (386 auf v. 280 auf; P < 0.05). These results suggest that BI embryos contain more and smaller LD than BT embryos and the LD described for BI embryo quality grade 1 are larger than those of quality grades 2 and 3, and even though the number of LD in morulae and blastocyst stage embryos are not different LD size is reduced as development occurs. Research funding provided by UNAM-DGAPA-PAPIIT IN200810.


2010 ◽  
Vol 22 (1) ◽  
pp. 248 ◽  
Author(s):  
J. H. F. Pontes ◽  
K. C. F. Silva ◽  
A. C. Basso ◽  
C. R. Ferreira ◽  
G. M. G. Santos ◽  
...  

In recent years, Brazil has become the leading country in the world for the number of embryos produced in vitro (Thibier M 2009 IETS Embryo Transfer Newsletter 22, 12-19). This is partly due to the large numbers of Bos indicus animals in Brazil, making up about 80% of the total cattle. The mean oocyte production per ultrasound-guided follicular aspiration from Bos indicus is higher than those for European breeds (Pontes JHF et al. 2009 Theriogenology 71, 690-697). In the present study, we analyzed 5407 ovum pick ups (OPU) and compared the average production of total (n = 90,086) and viable (n = 64,826) oocytes and the number of embryos produced in vitro from Gir (Bos taurus indicus), Holstein (Bos taurus taurus), 1/4 Holstein × 3/4 Gir, and 1/2 Holstein-Gir crossbreed cows. To obtain oocytes, OPU was repeated from 4 to 7 times (mean = 5.7 ± 2.4) in each donor cow aged from 3 to 7 years (mean = 5.0 ± 2.3) during a 12-mo period. COCs (n = 90,086) obtained were classified according to the presence of cumulus cells and the oocyte cytoplasm aspect (homogeneous or heterogeneous/fragmented). The viable oocytes (n = 64,826) were in vitro matured for 24 h at 38.8°C in an atmosphere of 5% CO2 in air. Since this was a commercial programm, frozen sexed semen (2 × 106 mL-1) from Gir (n = 8) or Holstein (n = 7) sires previously tested for high efficiency was used for IVF. Fertilization was carried out (18-20 h) and the presumed embryos were cultured for 7 days in the same conditions as were used for IVM. Data were analyzed by ANOVA. On average, 16.7 ± 6.2 oocytes were obtained per OPU/IVF procedure and 71.96% were considered viable. The mean numbers of total oocytes per OPU/IVF procedure were 17.1 ± 4.4 for Gir cows (n = 617), 11.4 ± 3.9 for Holstein cows (n = 180), 20.4 ± 5.8 for 1/4 Holstein × 3/4 Gir (n = 44), and 31.4 ± 5.6 for 1/2 Holstein-Gir crossbreed females (n = 37, P < 0.01). The mean numbers of viable oocytes per OPU/IVF procedure were 12.1 ± 3.8 for Gir cows, 8.0 ± 2.6 for Holstein cows, 16.8, ± 5.0 for 1/4 Holstein × 3/4 Gir, and 24.3 ± 4.7 for 1/2 Holstein-Gir crossbreed females (P < 0.01). The average number of embryos produced by OPU/IVF were 3.2 (n = 12,243/3378) for Gir cows, 2.2 (n = 2426/1138) for Holstein cows, 3.9 (n = 1033/267) for 1/4 Holstein × 3/4 Gir, and 5.5 (n = 1222/224) for 1/2 Holstein-Gir. The average number of embryos produced per IVF session from 1/2 taurus × indicus donor cows was greater (P < 0.01) than from Bos indicus cows. The number of recoverable and viable oocytes and the number of embryos produced in vitro from Bos indicus donors were higher than from Bos taurus females. Therefore, the highest oocyte yield and the greatest embryo production were obtained from 1/2 taurus × indicus females. This work was supported by In Vitro Brasil.


1991 ◽  
Vol 35 (1) ◽  
pp. 277 ◽  
Author(s):  
A. Subramaniam ◽  
K.P. Devarajan ◽  
M. Mohanan

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e12077
Author(s):  
Khalid Mahmood ◽  
Muhammad Zahid Tahir ◽  
Mahboob Ahmad Butt ◽  
Shazia Mansoor Qureshi ◽  
Amjad Riaz

Multiple Ovulation and Embryo Transfer (MOET) technology is a potential technique to upgrade livestock species’ genetics. The varied response to super-stimulatory treatments remains one of the limiting factors to this technology’s widespread use. The present study was aimed to improve the superovulation response and in-vivo embryo production by using controlled internal drug release (CIDR)-GnRH or CIDR-EB (Estradiol Benzoate) along with conventional superovulation protocol in Holstein Frisian (HF): Bos taurus; n = 42) and Crossbred (XB: Cholistani (Bos indicus) × HF; n = 28) cows. In the CIDR-GnRH/CIDR-EB treatment, CIDR was implanted in the cows after confirming the presence of a corpus luteum (CL) on the 8th day after estrus. 2 ml GnRH (Lecirelin acetate 0.0262 mg/ml) or 2 mg EB was also administered in CIDR-GnRH/CIDR-EB groups, respectively. Both groups were given super-stimulatory treatment from the 11th day after estrus (FSH in tapering doses twice a day for four consecutive days). On day 13, two doses of 2 ml prostaglandin (75 µg/ml of dextrorotatory cloprostenol) were administered (am: pm), and CIDR was removed the following day. Two artificial inseminations (AI) of the cows were performed (12 h apart) on the 15th day. No CIDR and GnRH/E.B were given in the control group, but the remaining superovulation protocol was the same. Later on, seven days after the first AI, non-surgical embryo flushing was done. The transferable embryos produced from three different superovulation protocols were then transferred into the recipient cows (n = 90) for determining their fertility. Statistical analysis revealed that the number of super-estrus follicles (SEF), multiple corpora lutea (MCL), ovulation/fertilization percentage, fertilized structures recovered (FSR), and transferable embryos (TEs) remained significantly higher (p < 0.05), and days taken for return to estrus (RTE) after embryo collection remained significantly lower (p < 0.05) in CIDR-GnRH (n = 18) and CIDR-EB (n = 15) groups as compared to the control (n = 37). The comparison between XB and HF cows revealed that the TEs production in CIDR-GnRH (XB = 5 vs HF = 13) and CIDR-EB (XB = 6 vs HF = 9) based superovulation protocols were 11.60  ±  4.08 vs 04.31  ±  0.98 and 09.33  ±  1.78 vs 05.22  ±  1.36, respectively. TEs production in XB cows (n = 5) of the CIDR-GnRH group was significantly higher (11.60  ±  4.08) than other groups. On the other hand, the days taken for RTE after embryo collection remained significantly lower (p < 0.05) in HF cows of treatment groups. However, the fertility of TEs was neither affected significantly (p > 0.05) by the superovulation protocol used nor by breed differences among donor cows. In conclusion, using CIDR-GnRH or CIDR-EB along with conventional superovulation protocol may enhance the efficiency of MOET programs in cattle. Furthermore, XB donor cows demonstrated a better performance than HF donor cows under subtropical conditions.


BMC Genomics ◽  
2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Cuili Pan ◽  
Zhaoxiong Lei ◽  
Shuzhe Wang ◽  
Xingping Wang ◽  
Dawei Wei ◽  
...  

Abstract Background Cyclin-dependent kinases (CDKs) are protein kinases regulating important cellular processes such as cell cycle and transcription. Many CDK genes also play a critical role during adipogenic differentiation, but the role of CDK gene family in regulating bovine adipocyte differentiation has not been studied. Therefore, the present study aims to characterize the CDK gene family in bovine and study their expression pattern during adipocyte differentiation. Results We performed a genome-wide analysis and identified a number of CDK genes in several bovine species. The CDK genes were classified into 8 subfamilies through phylogenetic analysis. We found that 25 bovine CDK genes were distributed in 16 different chromosomes. Collinearity analysis revealed that the CDK gene family in Bos taurus is homologous with Bos indicus, Hybrid-Bos taurus, Hybrid Bos indicus, Bos grunniens and Bubalus bubalis. Several CDK genes had higher expression levels in preadipocytes than in differentiated adipocytes, as shown by RNA-seq analysis and qPCR, suggesting a role in the growth of emerging lipid droplets. Conclusion In this research, 185 CDK genes were identified and grouped into eight distinct clades in Bovidae, showing extensively homology. Global expression analysis of different bovine tissues and specific expression analysis during adipocytes differentiation revealed CDK4, CDK7, CDK8, CDK9 and CDK14 may be involved in bovine adipocyte differentiation. The results provide a basis for further study to determine the roles of CDK gene family in regulating adipocyte differentiation, which is beneficial for beef quality improvement.


2021 ◽  
pp. 102998
Author(s):  
Bianca Vilela Pires ◽  
Nedenia Bonvino Stafuzza ◽  
Luara Afonso de Freitas ◽  
Maria Eugênia Zerlotti Mercadante ◽  
Ester Silveira Ramos ◽  
...  

Author(s):  
J Ranches ◽  
R Alves ◽  
M Vedovatto ◽  
E Anne Palmer ◽  
P Moriel ◽  
...  

Abstract A two-year study was conducted at the University of Florida – IFAS, Range Cattle Research and Education Center (Ona, FL) to evaluate differences in the metabolism of Cu and Se of Angus (Bos taurus) and Brahman (Bos indicus) cattle. Thirty-two pregnant beef cows (n = 8 Brahman and 8 Angus/year) were enrolled in the study in the first trimester of gestation. The study consisted of 3 phases: (1) restriction (d 0 to d 90); (2) supplementation (d 91 to 150), and (3) calving. During all 3 phases, cows were individually fed and housed in partially covered drylot pens. During the restriction and supplementation phases cows were provided a 1.5 kg/d of a grain-based concentrate supplement, which was fortified with flowers of S (50 g of supplemental S/cow daily; restriction phase) or Cu and Se (100 and 3 mg/d of Cu and Se, respectively; supplementation phase). Blood and liver samples were collected from all cows on 30 d intervals and from both cows and calves within 24 h of calving. Colostrum and milk samples were collected at calving and 7 d after birth. All data were analyzed using the MIXED procedure of SAS, where cow and calf were the experimental unit. During the restriction phase, a breed × day effect (P = 0.03) was observed where Brahman had greater liver Cu concentration than Angus cows in all sampling days. For liver Se concentration, a tendency (P = 0.07) for a breed effect was observed where Angus cows tended to have greater liver Se concentration than Brahman. During the supplementation phase, breed (P &lt; 0.001) and day (P &lt; 0.01) effects were observed, where Brahman cows had greater liver Cu concentration than Angus. For liver Se concentration, a day effect (P &lt; 0.001) was observed, where liver Se concentration increased (P &lt; 0.001) from d 90 to 120 and remained unchanged (P = 0.86) until d 150. At calving no effects of breed (P = 0.34) were observed for liver Cu concentration of cows, however, Brahman calves tended (P = 0.09) to have greater liver Cu concentration than Angus calves. For Se liver concentration at calving, Angus cows tended (P = 0.07) to have greater liver Se concentration than Brahman cows, however no breed differences (P = 0.70) were observed for liver Se concentration of calves at birth. In summary substantial differences in multiple indicators of Cu and Se status were observed between Angus and Brahman cattle, implying that Angus and Brahman cattle possibly have different mechanisms to maintain adequate Cu and Se status.


2011 ◽  
Vol 89 (9) ◽  
pp. 2859-2866 ◽  
Author(s):  
S. F. M. Bonilha ◽  
L. O. Tedeschi ◽  
I. U. Packer ◽  
A. G. Razook ◽  
R. F. Nardon ◽  
...  

1982 ◽  
Vol 14 (1) ◽  
pp. 1 ◽  
Author(s):  
CRE Halnan ◽  
Janine I Watson
Keyword(s):  

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