246 INFLUENCE OF BREED AND SEASON ON IN VITRO EMBRYO PRODUCTION

2016 ◽  
Vol 28 (2) ◽  
pp. 255
Author(s):  
B. Bernal ◽  
J. Revol ◽  
J. M. Oviedo ◽  
A. Tribulo ◽  
H. Tribulo ◽  
...  
Keyword(s):  
Bos Taurus ◽  
Bos Indicus ◽  
In Vitro Production ◽  
Embryo Production ◽  
Significant Difference ◽  
In Vitro Embryo Production ◽  
Follicle Aspiration ◽  
Vitro Production ◽  
Difference Test

A retrospective analysis of in vitro production (IVP) data was done to determine the influence of breed and season on the production of viable oocytes and embryos. Cumulus‐oocyte complexes (COC) were obtained from 1946 ultrasound-guided follicle aspiration (ovum pickup) sessions performed at random stages of the oestrous cycle without superstimulation in Bos taurus and Bos indicus donors in commercial IVP in Argentina. Frozen-thawed conventional semen was used in beef cattle and conventional (n = 139) and sexed-selected (n = 481) semen in dairy cattle. The COC were classified, matured in B-199 medium, fertilized in IVF-SOF medium (Day 0), and cultured in SOF medium supplemented with 0.4% BSA under oil at 38.8°C, 5.5% CO2, and saturated humidity for 7 days. The number of viable COC and transferable embryos in each breed and season were compared by ANOVA and means were compared by Fisher’s Least Significant Difference test. Proportions were first transformed by arcsin and then analysed by ANOVA. To simplify the interpretation of the results, breeds were grouped as follows: dairy Bos taurus (Holstein, n = 620), beef Bos taurus (Angus and Bonsmara, n = 229), Bos taurus × Bos indicus (Brangus and Braford, n = 1045), and Bos indicus (Brahman, n = 52). There was no interaction between breed and season for any of the end points analysed (P > 0.1). Mean (± standard error of the mean) numbers of viable COC and transferable embryos were higher (P < 0.01) in Bos indicus × Bos taurus (19.3 ± 0.4 and 5.3 ± 0.2, respectively) and Bos indicus (15.8 ± 1.4 and 6.8 ± 0.9, respectively) than in beef (11.6 ± 0.5 and 3.0 ± 0.2, respectively) and dairy (8.0 ± 0.2 and 1.6 ± 0.1, respectively) Bos taurus donors. Cleavage rates were higher (P < 0.01) in Bos indicus (72%) than in the other breeds (57% for Bos indicus × Bos taurus and dairy Bos taurus and 54% for beef). Transferable embryo rates were higher (P < 0.01) in Bos indicus (41%) and Bos indicus × Bos taurus (30%) than in beef Bos taurus (26%). Dairy Bos taurus had the lowest (P < 0.01) embryo rates of all breeds (21%). In dairy Bos taurus, cleavage rates, the number of embryos produced, and transferable embryo production rates were higher (P < 0.01) when conventional semen was used (62%, 2.8 ± 0.15, and 27%, respectively) compared to sexed-selected semen (55%, 1.3 ± 0.1, and 19%, respectively). With regards to season, the number of viable COC was highest (P < 0.01) in the spring (14.3 ± 0.5), lowest in the summer (11.3 ± 1.0), and intermediate in the fall (12.2 ± 1.2) and winter (13.7 ± 1.2), which did not differ. Although not affected significantly by season, the number of embryos produced was numerically lower in the summer (2.8 ± 0.4) than in the spring (4.2 ± 0.2), winter (4.5 ± 0.5), or fall (4.6 ± 0.5). In conclusion, in vitro embryo production was directly influenced by breed and season. Bos indicus influenced cattle and the spring season were preferable for commercial IVP programs that did not include superstimulation.

scholarly journals In vitro performance of Zebu (Bos indicus) and Taurus (Bos taurus) donor cow embryos

2020 ◽  
Vol 21 ◽  
Author(s):  
Anita Soares Barbosa GUIMARÃES ◽  
Laiara Fernandes ROCHA ◽  
Ronival Dias Lima de JESUS ◽  
Gisvani Lopes VASCONCELOS ◽  
Gabriela ANGHINONI ◽  
...  
Keyword(s):  
Culture Medium ◽  
Bos Taurus ◽  
Bos Indicus ◽  
In Vitro Production ◽  
Embryo Production ◽  
Maturation Medium ◽  
Significant Difference ◽  
Vitro Production ◽  
In Vitro Performance

ABSTRACT In this study, the in vitro production of bovine embryos from zebu and taurine donors was compared. Cumulus-oocyte complexes (COCs) were obtained from 167 Bos taurus and 161 Bos indicus donors by ovum pick-up. COCs were classified based on their morphological quality, matured in incubators for 22 to 24 h in maturation medium, and then fertilized for 18 to 22 h. The zygotes were transferred to the culture medium for seven days. The embryos were classified as morula (OM), initial blastocyst (BI), blastocyst (BL), and expanded blastocyst (BX), before being transferred to synchronized recipient cows. Pregnancy was diagnosed 30-45 days post-transfer. The Bos indicus donors had a higher oocyte yield (n = 2556) than Bos taurus donors (n = 1903) (P = 0.008). The COCs from zebu donors had a better morphological quality than those from taurine donors (n = 689 vs. 444 for grade 1 COC, P < 0.0001; n = 681 vs. 509 for grade 2 COC, P = 0.010, for zebu and taurine donors, respectively). There were differences in embryo production percentages obtained from OM (0.44% from zebu and 6.42% from taurine, P = 0.017), BL (14.18% from zebu and 3.74% from taurine, P < 0.0001), and BX (81.43% from zebu and 75.13% from taurine, P < 0.0001). No significant difference was observed for embryo production from BI and pregnancy rate (P > 0.05). The Bos indicus cows showed greater oocyte recovery, number of viable oocytes, and production of viable embryos than the Bos taurus cows.

257 DIFFERENCES BETWEEN BOS TAURUS AND BOS INDICUS EMBRYOS: TRANSCRIPTS AMOUNTS

2010 ◽  
Vol 22 (1) ◽  
pp. 285
Author(s):  
S. Wohlres-Viana ◽  
M. M. Pereira ◽  
A. P. Oliveira ◽  
J. H. M. Viana ◽  
M. A. Machado ◽  
...  
Keyword(s):  
Production System ◽  
Production Systems ◽  
Bos Taurus ◽  
Bos Indicus ◽  
In Vitro Production ◽  
Embryo Production ◽  
Peroxiredoxin 1 ◽  
Vitro Production

The Zebu breeds (Bos indicus) are different from European breeds (Bos taurus) in some aspects of their reproductive physiology, including follicle recruitment, number of follicular waves, and oocyte ultrastructure. On the other hand, embryos produced in vivo and in vitro show morphological and developmental differences, which can be related to culture environment. The aim of this study was to evaluate the effect of breed (Gyr v. Holstein) within embryo production system (in vivo and in vitro), as well as effect of production systems within breeds on relative abundance of transcripts related to formation, survival, and subsequent development of blastocysts, such as those involved in water and small solutes transport (Aquaporins 3 and 11), blastocoel formation (Na+/K+-ATPase a1 and |52), and cellular stress response (Peroxiredoxin 1). For in vivo embryo production, donors were superstimulated with FSH and inseminated, and embryos were recovered 7 days after AI. For in vitro embryo production, oocytes recovered by ovum pickup were in vitro matured and fertilized and then cultured for 7 days in culture medium under 5% CO2 at 38.5°C. For each group, blastocysts (n = 15) distributed in 3 pools were used for RNA extraction (RNeasy MicroKit, Qiagen, Valencia, CA, USA), followed by RNA amplification (Messageamp II amplification kit, Ambion-Applied Biosystems, Foster City, CA, USA) and reverse transcription (SuperScript III First-Stand Synthesis Supermix, Invitrogen, Carlsbad, CA, USA). The cDNA were submitted to real-time PCR, using the H2a gene as endogenous control, and analyzed by REST© software. To evaluate breed effect within the production systems, 2 comparisons were performed: (1) in vivo: Gyr v. Holstein and (2) in vitro: Gyr v. Holstein, considering Holstein data as 1.00. To evaluate production system effect within breeds, 2 comparisons were performed: (1) Gyr: in vivo v. in vitro and (2) Holstein: in vivo v. in vitro, considering in vivo produced embryo data as 1.00. The results are shown as mean ± SEM. For in vivo comparison between breeds, Aquaporin 3 (1.66 ± 0.77), Na+/K+-ATPase a1 (1.61 ± 0.56), and Peroxiredoxin 1 (1.61 ± 0.66) were up-regulated (P < 0.05) in Gyr embryos when compared with Holstein embryos, whereas for in vitro comparison, no differences (P > 0.05) were found. For comparisons between production systems within breeds, only Peroxiredoxin 1 (0.31 ± 0.39) was down-regulated (P < 0.01) in in vitro produced Gyr embryos when compared with in vivo counterparts. No differences (P > 0.05) were found between production systems for the Holstein breed. In conclusion, these data suggest that there is a difference on gene expression between Bos taurus and Bos indicus blastocysts, but such difference between breeds can be attenuated by the in vitro production system, indicating an embryo adaptation to the in vitro culture conditions. The data also suggest that the in vitro production system can influence the amount of transcripts in Gyr embryos. Other genes should be evaluated for a better understanding of these differences. Financial support was provided by CNPq and FAPEMIG.

scholarly journals Effects of a high-energy diet on oocyte quality and in vitro embryo production in Bos indicus and Bos taurus cows

2015 ◽  
Vol 98 (5) ◽  
pp. 3086-3099 ◽  
Author(s):  
J.N.S. Sales ◽  
L.T. Iguma ◽  
R.I.T.P. Batista ◽  
C.C.R. Quintão ◽  
M.A.S. Gama ◽  
...  
Keyword(s):  
Bos Taurus ◽  
Bos Indicus ◽  
Oocyte Quality ◽  
High Energy ◽  
Embryo Production ◽  
In Vitro Embryo Production

scholarly journals Synchronization with estradiol benzoate in the presence of the corpus luteum in Wagyu cows increases the number of medium follicles but does not interfere with in vitro production of embryos

2021 ◽  
Vol 42 (3) ◽  
pp. 1147-1158
Author(s):  
Maria Fernanda Zamai ◽  
◽  
Fábio Luiz Bim Cavalieri ◽  
Marcia Aparecida Andreazzi ◽  
Fabio Morotti ◽  
...  
Keyword(s):  
Corpus Luteum ◽  
Estradiol Benzoate ◽  
Breeding Systems ◽  
In Vitro Production ◽  
Embryo Production ◽  
Follicular Wave ◽  
In Vitro Embryo Production ◽  
Oocyte Donors ◽  
Vitro Production

Reproductive biotechnologies are emerging as an important element for livestock; however, some strategies must be modified to adapt to different breeding systems, such as the use of follicular synchronization protocols. This study aimed to evaluate follicular synchronization using estradiol benzoate (EB), in the presence of the corpus luteum (CL) from Wagyu oocyte donors in in vitro embryo production (IVEP). Rounds of IVEP were performed in heifers and cows (n=19) that were classified into three groups: G1/CL - animals with CL, G2/WCL - animals without CL, and G3/CL + EB - animals with CL that were subjected to follicular synchronization with EB at D0. The groups G1/CL and G2/WCL were considered the control and undertook the natural process of follicular dynamics. The results showed that the synchronization of the follicular wave with the application of EB in the presence of CL, presented a smaller number of small (6.05 ± 0.55) and large follicles (0.45 ± 0.15), but increased (P < 0.05) the number of medium-sized follicles (16.20 ± 0.90). However, the results of ovum pick up showed that regardless of whether or not EB was applied, and regardless of the presence or absence of CL in the Wagyu donor, there was no difference among the groups (P > 0.05) concerning the number of viable oocytes and the viability rate. It was concluded that follicular synchronization using EB in Wagyu oocyte donors that presented a CL, increased the number of medium-sized follicles. However, there was no improvement in the efficiency of ovum pick up, in vitro embryo production, and pregnancy rate.

86 Comparison of in vivo and in vitro embryo production in Bonsmara donors

2019 ◽  
Vol 31 (1) ◽  
pp. 168
Author(s):  
B. H. Bernal ◽  
J. L. Barajas ◽  
J. A. Ortega ◽  
A. Cedeño ◽  
S. Andrada ◽  
...  
Keyword(s):  
Bos Taurus ◽  
Gonadotropin Releasing Hormone ◽  
P Value ◽  
In Vitro Production ◽  
Embryo Production ◽  
Releasing Hormone ◽  
Gradient Technique ◽  
Vitro Production

A retrospective analysis of embryo production records from 2013 to 2017 was carried out to evaluate the in vivo and in vitro production (IVP) of embryos in donors of the Bonsmara breed (i.e. tropically adapted Bos taurus). Only donors with production records of both in vivo and in vitro embryos during the same period were used. A total of 127 superovulations and ova/embryo collections of 19 donors were evaluated. The donors were superstimulated with the following protocol: on Day 0 they received a device with 1g of progesterone (DIB, Zoetis, Argentina), 50mg of rogesterone (Progestar, Zoetis), and 5mg of oestradiol-17β (17ßOestradiol, Rio de Janeiro, Argentina) or 2mg of oestradiol benzoate (Gonadiol, Zoetis) intramuscularly (IM) at the same time. Superstimulatory treatments were initiated on the morning of Day 4 with Folltropin-V (Vetoquinol, France; total dose=240 to 340mg IM) in twice-daily decreasing doses over 4 days. All donors received 2 IM injections of 500µg of cloprostenol (Ciclase DL, Zoetis) on the morning and afternoon of Day 6 and; the intravaginal devices were removed on the morning of Day 7 and 100µg of Gonadorelin (gonadotropin-releasing hormone, Gonasyn gdr, Zoetis) was given on the morning of Day 8. Donors were inseminated using semen from 9 Bonsmara bulls, 12 and 24h after gonadotropin-releasing hormone. On Day 15, ova/embryos were collected and classified according to IETS standards. A total of 89 follicular aspirations (ovum pickup) of 19 donors for IVP were evaluated. The ovum pickups were performed at random stages of oestrous cycle, without superstimulation or other hormone treatments. A total of 1109 viable oocytes (12.5±0.9 per ovum pickup) were collected and matured for 24h in 100-µL drops of maturation medium (TCM-199, supplemented with hormones) under mineral oil and incubated at 38.5°C in 5.5% CO2 and humidity at saturation. Fertilization was performed using 3 Bonsmara bulls that were also used for in vivo embryo production. Viable sperm were obtained using the percoll gradient technique (45-90%). The sperm pellet was dissolved in TL-Sperm, centrifuged, and then diluted to a final concentration of 1.5×106 sperm/mL. Zygotes were stripped and placed in drops of 100µL of SOF medium supplemented with 0.4% BSA under oil at 38.8°C, 5.5% CO2, 7% O2, and humidity at saturation for 7 days. The culture medium was renewed on Days 3 and 5. The data were analysed using the GLM procedure of SAS (SAS Institute Inc., Cary, NC, USA), a P-value &lt;0.05 was considered significant. The mean (±standard error of the means) number of CL, ova/embryos collected, fertilized ova, and transferable embryos were 12.9±0.6, 8.8±0.6, 6.6±0.5, and 4.7±0.4, respectively. A total of 662 oocytes (66.3±2.4%) cleaved 48h post-IVF. On Day 7, an average of 4.4±0.3 embryos were produced. No differences were detected in the number of transferable embryos produced in vivo v. those produced in vitro. Furthermore, no significant differences were found between the techniques or bulls on the proportion of embryos produced in relation to the ova/embryos or oocytes obtained (in vivo 51.5±3.2% v. in vitro 42.9±2.5%). In conclusion, the in vivo and in vitro production of embryos are both effective alternatives to increase the number of offspring from valuable Bonsmara donors.

130 In vitro embryo production using frozen semen from cloned and non-cloned Bos indicus bulls

2019 ◽  
Vol 31 (1) ◽  
pp. 190
Author(s):  
S. Romo ◽  
O. Sebastián ◽  
F. Guerrero ◽  
R. Romero ◽  
F. Muñoz ◽  
...  
Keyword(s):  
Bos Indicus ◽  
The State ◽  
Assisted Reproductive Techniques ◽  
Embryo Production ◽  
Humid Atmosphere ◽  
Frozen Semen ◽  
Significant Difference ◽  
Reproductive Techniques ◽  
In Vitro Embryo Production

Reproductive biotechnology has continued to evolve rapidly, allowing the development of techniques to increase reproductive efficiency and contribute to the genetic improvement of cattle. Some of these techniques include the in vitro maturation and IVF of oocytes, sperm sexing and cloning. These modern assisted reproductive techniques can help produce offspring of desired genetic characteristics and of a pre-determined sex. However, studies of the bull’s contribution to in vitro reproductive performance are scarce in the Brahman breed. The aim of this study was to compare oocyte maturation and embryo production in vitro using frozen semen from 5 Brahman bulls (Bos indicus), cloned (n=1) and non-cloned (n=4), with characteristics and genetics of high commercial value. The age of the bulls at the time of semen collection and cryopreservation ranged from 2 to 7 years. The oocytes were obtained on 2 different dates (45 days between collections) using pooled oocytes collected by ovum pickup at random stages of the oestrous cycle, from a total of 15 Brahman donor cows. Oocytes were transported to a laboratory in the State of Chiapas, Mexico (Genemex Internacional). The oocytes were cultured in maturation medium for 24h. For IVF, conventional semen was used from one bull (B1) and his clone (B12), the grandson of B1 (B2), and 2 non-related bulls (B3 and B4). The gametes were co-incubated for 22h and afterward placed in medium for embryo development and cultured for 7 days in a humid atmosphere with 5% CO2 in air. Of the matured oocytes, 36/43 (84%), 16/32 (50%), 101/143 (70%), 46/67 (68%) and 53/65 (81%) were fertilized using semen from B1, B12, B2, B3 and B4, respectively. Of the fertilized oocytes, 15/30 (50%), 8/16 (50%), 45/101 (44%), 21/46 (45%) and 18/53 (34%) resulted in transferrable embryos, corresponding to semen from the same bulls, respectively. This would appear to be the first scientific report in Mexico about the use of semen from a cloned bull for in vitro embryo production. In IVF, similar results were observed between B1 and a non-related bull (B4). Similar results in transferrable embryos were observed between B1 and B12 but also similar to a related bull (B3) and a non-related bull (B4). A Fisher’s exact test of the IVF results comparing B1 and B12 found a significantly (P&lt;0.05) higher number of fertilized oocytes for B1. However, a significant difference was not found (P&gt;0.05) concerning the number of transferrable embryos produced by these two bulls. In conclusion, the Brahman bulls in this study differ in their contribution to IVF and embryo production. Further studies are required to determine the factors responsible for such effects, e.g. age differences or clone versus non-clone mosaicism. Results from this research contribute to the study and development of assisted reproductive techniques for increasing in vitro production efficiency in Zebu cattle. We thank the Rosales family, from El Herradero Ranch, in the State of Campeche, Mexico, for allowing the use of their cattle for this project.

scholarly journals Effect of the antral follicle count of Bos taurus × Bos indicus dairy cows on in vitro embryo production

2020 ◽  
pp. 2171-2178
Author(s):  
Sheila Merlo Garcia ◽  
Paula Alvares Lunardelli ◽  
Kleber Luciano Ancioto ◽  
Eduardo Cardoso de Oliveira ◽  
Larissa Zamparone Bergamo ◽  
...  
Keyword(s):  
Bos Taurus ◽  
Bos Indicus ◽  
Antral Follicle ◽  
Estradiol Benzoate ◽  
Antral Follicle Count ◽  
Success Rates ◽  
In Vitro Production ◽  
Follicular Aspiration ◽  
Vitro Production

This study aimed to evaluate the potential of Girolando (Bos taurus × Bos indicus) cows with high and low antral follicle counts (AFC) for the in vitro production of bovine embryos (IVEP), as well as the pregnancy rates of the recipients of these embryos. Girolando cows (Bos taurus × Bos indicus) were classified as high and low AFC when they had 35-52 (n = 13) and 11-17 follicles (n = 15), respectively. All animals were subjected to repeated follicular aspiration [Ovum pick-up (OPU)] and subsequent IVEP sessions. The synchronization protocol of the recipients was performed on a random day of the estrous cycle (Day 0) with the implantation of progesterone, estradiol benzoate, and prostaglandin. The high AFC group had higher aspirated oocyte/OPU (42.6 ± 5.2 vs. 14.6 ± 1.9; p < 0.01) and cultured oocyte/OPU (38.1 ± 6.6 vs. 12.3 ± 2.8; p < 0.01) averages as well as a higher blastocyst percentage on D7 (23.0 ± 1.0% vs. 18.4 ± 1.5%; p < 0.05) and higher pregnancy rate (42.7 ± 2.7% vs. 39.7 ± 4.6%; p < 0.05) than the low AFC group. Thus, we can conclude that animals with high AFC had better IVEP success rates than animals with low AFC.

scholarly journals Effect of quality of sperm chromatin structure on in -vitro production of cattle embryos

2005 ◽  
Vol 48 (1) ◽  
pp. 32-39
Author(s):  
L. Kątska-Książkiewicz ◽  
M. Bochenek ◽  
B. Ryńska
Keyword(s):  
Chromatin Structure ◽  
Blastocyst Stage ◽  
Sperm Chromatin ◽  
In Vitro Production ◽  
Embryo Production ◽  
In Vitro Embryo Production ◽  
Bull Ejaculate ◽  
Vitro Production

Abstract. Bull effect on results of in vitro embryo production has been well documented. The aim of the present study was to find the relationship between quality of bull sperm chromatin and its effect on in vitro embryo production. Bovine in vitro matured oocytes were fertilized in vitro using capacitated spermatozoa (freshly ejaculated or frozen-thawed) of 12 bulls. Semen was simultaneously processed according to the sperm chromatin structure assay (SCSA) method and was analysed by flow cytometry. At least 3 replications of IVP with the same semen sample were done. The percentage of spermatozoa with abnormal chromatin ranged from 0.4% to 23.8%. All bulls used for the experiment were divided into three groups showing minimal (0.82% ± 6.82%), low (1.70% ± 15.82%) and high (18.16% ± 53.59%) percentages of spermatozoa with abnormal chromatin structure. Both cleavage rates and embryo development to the blastocyst stage were correlated significantly with sperm chromatin abnormalities and resulted in 23.1, 17.7 and 12.2% of blastocysts respectively for sperm with minimal, low and high percentages of chromatin abnormalities. The SCSA method may be used as a practical indicator of suitability of bull ejaculate for IVP purposes.

222 COMPARISON OF COMMERCIAL IN VITRO EMBRYO PRODUCTION OF BRAHMAN DONORS UNDER BRAZILIAN v. PANAMANIAN MANAGEMENT

2011 ◽  
Vol 23 (1) ◽  
pp. 210
Author(s):  
J. R. S. Chen ◽  
L. F. Nasser ◽  
L. Penteado ◽  
M. Mendizabal ◽  
A. C. Basso ◽  
...  
Keyword(s):  
Large Scale ◽  
Bos Taurus ◽  
Bovine Embryo ◽  
In Vitro Production ◽  
Embryo Production ◽  
Test Analysis ◽  
Animal Biotechnology ◽  
Follicular Aspiration ◽  
In Vitro Embryo Production

Brazil is a leading country in the world in commercial use of in vitro-produced bovine embryos, with approximately 200 000 transfers per year (IETS; Thibier, 2009). This model of large-scale commercial in vitro bovine embryo production is now available for Panamanian producers. Because of the tropical environment in Panama, the most popular breed is the Brahman, a Zebu type of cattle that has been shown to have more follicles emerging per follicular wave than Bos taurus type of cattle and, consequently, that produce more oocytes per session of follicular aspiration. This characteristic, added to the embryo production results, permits such a biotechnology to be implemented on a commercial scale and incorporated into the reproduction management of a herd. A comparison of oocyte number and quality, cleavage, and embryo production was made using the same in vitro production system (InVitro Brazil, Mogi Mirim, São Paulo, Brazil) for Brahman donors, both in Brazil and in Panama. Data were compared using a z-test analysis (Table 1). The percentage of cleaved zygotes was greater (P < 0.001) with the Panamanian Brahman donors as compared with the Brazilian Brahman donors (73 v. 69%, respectively). However, the percentage of blastocysts/cleaved zygotes was greater (P < 0.01), indicating a higher blastocyst production rate from the Brazilian donors. No other differences were observed. Thus, in vitro embryo production with Brahman donors could be used as a tool to improve and spread superior genetics within a Panamanian herd and could also serve as a model for other Central American and Caribbean countries under similar management systems. Table 1.Panama and Brazil in vitro Brahman embryo production1 This work was supported by BORN Animal Biotechnology, Panama City, Panama.

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