132 TIMING OF THE FIRST ZYGOTIC CLEAVAGE WAS NOT RELATED TO THE SHIFT IN SEX RATIO OF BOVINE BLASTOCYST IN VITRO

2009 ◽  
Vol 21 (1) ◽  
pp. 165
Author(s):  
K. Zaorska ◽  
E. Pers-Kamczyc ◽  
D. Lechniak
Keyword(s):  
Sex Ratio ◽  
Positive Influence ◽  
Control Group ◽  
Fish Analysis ◽  
Chi Square ◽  
Experimental Conditions ◽  
Female Ratio ◽  
Developmental Potential ◽  
Frozen Semen

It has been shown that embryos which cleaved earlier (<30 h postinsemination, hpi) have greater chance to develop to blastocyst. It has been shown that in in vitro conditions male embryos develop faster than females. Further, spermatozoa with the Y chromosome more frequently penetrate oocytes during the first 6 hpi. The aim of this experiment was to investigate the sex-related embryo growth rate in relation to the timing of the first zygotic cleavage and GH presence during IVM. Oocytes were matured in TCM-199 supplemented with fatty acid free BSA and hormones (FSH and GH) and then inseminated (Parrish et al. 1998 Biol. Reprod. 38, 1171–1180), whereas embryos were cultured in sequential media (Lane et al. 2005 Theriogenology 60, 407–419). All embryos that cleaved by 30 hpi (early cleavers, EC) were selected and cultured separately. The remaining embryos cleaved by 48 hpi (non early cleavers, NEC) were also incubated in separate drops. Blastocysts of proper morphology were collected at 176 hpi and subjected to sex determination by PCR (AMGL gene). The significance of developmental stage, timing of the first zygotic cleavage and GH supplementation in relation to the sex ratio was evaluated by the chi-square test. All straws with frozen semen of 2 bulls used in this experiment were derived from single ejaculates. The sex ratio of sperm samples used for IVF was evaluated by FISH with locus-specific probes. The experiment was done on 266 embryos obtained from 1249 oocytes. A significant predominance of male blastocysts regardless of the experimental conditions was observed [the male to female ratio (M:F) 2:1, P < 0.001]. FISH analysis revealed that there was no deviation from the expected 1:1 ratio of X and Y spermatozoa in sperm samples used for IVF. When the timing of the first zygotic cleavage was considered, shift in M:F ratio in favor of males (P < 0.01) in blastocysts derived from EC zygotes was noticed. Due to a very low number of NEC embryos, this category was not included in analysis. Although the M:F ratio was shifted towards males, the rate of female embryos was greater in the control group (25F/38M) v. GH group (16F/39M) of EC expanded blastocysts. This phenomenon was not observed among hatched blastocysts; however, GH presence caused an increase (P < 0.01) in the number of females at this stage. Therefore GH may stimulate embryonic growth, especially embryos of reduced quality, through its positive influence on cytoplasmic oocyte maturation. In conclusion, the predominance of male blastocysts observed in this study may be attributed to the applied IVF procedure because the X:Y ratio in spermatozoa was not different from the expected 1:1 ratio. Moreover, significantly fewer females among analyzed blastocysts may suggest that the developmental potential of female embryos in applied in vitro conditions was somehow reduced when compared with males. This became evident when the transition from expanded to hatched blastocysts was observed. This work was supported by the project No. N302 046 31/3780 of the Ministry of Science and Higher Education, Poland.

198 Comparison of invitro production of bison and cattle embryos and effect of L-carnitine during maturation of bison oocytes

2020 ◽  
Vol 32 (2) ◽  
pp. 227
Author(s):  
A. R. Moawad ◽  
H. Benham ◽  
J. P. Barfield
Keyword(s):  
Assisted Reproductive Technologies ◽  
Cell Mass ◽  
Reproductive Technologies ◽  
T Test ◽  
Control Group ◽  
Chi Square ◽  
Important Species ◽  
Developmental Potential ◽  
Cell Numbers ◽  
Frozen Semen

Bison are an important species in North America, both economically and culturally. Although assisted reproductive technologies have been applied to preserve the genetic diversity of bison, development of these technologies remains limited for this species. The objective of the present study was to compare success rates of oocyte maturation, fertilization, and embryo development invitro in bison versus cattle (experiment 1). Cumulus-oocyte complexes obtained from abattoir-derived cattle and bison ovaries were matured, fertilized with frozen semen, and cultured invitro using standard procedures (De La Torre-Sanchez et al. 2006 Reprod. Fertil. Dev. 18, 585-596). At least three replicates were repeated for each experimental group. Oocyte recovery rate was lower in bison than in cattle (4.3; 2797/699 vs. 6.7; 4138/677, oocyte/ovary; P&lt;0.01, t-test). Nuclear maturation (oocytes at MII, 23h post-IVM) and fertilization rates (oocytes with 2 pronuclei 18h post-insemination; p.i.) evaluated by Hoechst stain were lower (P&lt;0.01, chi-square) for bison (65.1%; 56/86 and 32.7%; 18/55, respectively) than for cattle (88.3%; 83/94 and 70.9%; 39/55, respectively). Polyspermy tended to be higher in bison than in cattle (12.7% vs. 3.6%, P=0.08). The percentages of 2-cell embryos tended to be lower in bison than in cattle (13.5% vs. 25.0%, P&gt;0.05) at 24h p.i. but by 30h p.i., this difference increased (33.7% vs. 67.0%, P&lt;0.01, chi-square). Cleavage (Day 3) and blastocyst (Day 7) rates were lower (P&lt;0.01, chi-square) for bison (58.2%; 280/481 and 14.6%; 70/481, respectively) than for cattle (90.8%; 405/446 and 22.9%; 102/446, respectively). Total cell number (74.9±4.8 vs.114.2±5.8), trophectoderm cell numbers (57.9±4.6 vs. 89.2±4.8) and inner cell mass cell numbers (16.9±2.3 vs. 25±1.9) as determined by Hoechst and propidium iodide were all lower (P&lt;0.01, t-test) in bison than in cattle blastocysts. To improve oocyte competence in bison, we evaluated effects of L-carnitine (LC) supplementation during IVM on developmental potential of bison oocytes (experiment 2). Cumulus-oocyte complexes were matured in IVM medium supplemented with 0, 0.15, 0.3, 0.6, or 1.2mgmL−1 LC. No differences were observed in cleavage rates of control (0mgmL−1 LC) and LC-treated groups (values ranged from 60.0 to 66.4%). Interestingly, a dose-dependent increase in blastocyst development was found with the lowest value recorded in control group (10.4%; 14/134) and the highest value in the 1.2mgmL−1 LC supplemented group (22.2%; 23/105; P&lt;0.01, chi-square, n=4). Adding 1.2mgmL−1 LC to the IVM medium improved the percentage of hatching blastocysts compared with the control. In conclusion, bison oocytes exhibited lower invitro maturation, fertilization, and developmental rates compared with cattle oocytes using our system, and bison embryos were delayed in the timing of first cleavage. L-Carnitine supplementation during IVM of bison oocytes improved the preimplantation development and quality of invitro-produced blastocysts.

43 SHORT- AND LONG-LASTING EFFECTS OF TRICHOSTATIN A TREATMENT OF SCNT EMBRYOS IN CATTLE

2011 ◽  
Vol 23 (1) ◽  
pp. 127 ◽  
Author(s):  
I. Lagutina ◽  
R. Duchi ◽  
S. Colleoni ◽  
G. Lazzari ◽  
C. Galli
Keyword(s):  
Trichostatin A ◽  
Blastocyst Stage ◽  
Development Project ◽  
Full Term ◽  
Control Group ◽  
Cloning Efficiency ◽  
Chi Square ◽  
Developmental Potential ◽  
Student’S T

Both preimplantation and full-term development of mouse somatic cell nuclear transfer (SCNT) embryos are significantly enhanced by treatment with trichostatin A (TSA), an inhibitor of histone deacetylase (Kishigami et al. 2006 Biochem. Biophys. Res. Commun. 340, 183–189; Rybouchkin et al. 2006 Biol. Reprod. 74, 1083–1089). The present study was designed to examine the effect of TSA treatment on preimplantation and full-term development of bovine cloned embryos. To investigate the effect of TSA on bovine NT embryos development, we treated them with 50 nM TSA during the first 10 h after activation. Bovine NT-embryos were reconstructed using adult fibroblasts of 2 female donors (A and B) with significantly different in vitro cloning efficiency (respectively, 84/245; 34.3% v. 155/298; 52.1% blastocyst D7, P ≤ 0.05, chi-square test). TSA treatment significantly improved blastocyst rate in A, however did not affect development in B (56.3% and 50.5%, respectively). The level of acetylated histone H3K9 10 h after activation detected by anti-acH3K9 antibody was significantly increased after TSA-treatment in A (P ≤ 0.05, Student’s t-test) but did not change in B, thus demonstrating that the levels of histone acetylation in cloned embryos correlate with their in vitro developmental potential. To evaluate the long-lasting effect of TSA-treatment on the full-term development of cloned embryos, SCNT embryos derived from 4 female donor animals were reconstructed. 196 TSA-treated embryos at the blastocyst stage were transferred into 98 recipients and 2 calves (2%) were born. In the control group, 167 embryos were transferred into 141 recipients and 3 calves (2.1%) were born. Our data show that cell lines demonstrate different susceptibility to TSA that may affect reprogramming of the somatic genome with low level of acetylation resulting in higher in vitro embryo development. However, TSA does not improve overall cloning efficiency in cattle, measured as full-term development. Project partly supported by EU grants Plurisys (n 22348), Xenome (LSHB-CT-2006-037377) and Regione Lombardia.

296 FREQUENT RECTAL PALPATIONS FOLLOWING SUPEROVULATORY TREATMENT AFFECT SEX RATIO OF EMBRYOS RECOVERED FROM HOLSTEIN HEIFERS

2008 ◽  
Vol 20 (1) ◽  
pp. 228
Author(s):  
A. Ideta ◽  
K. Hayama ◽  
M. Urakawa ◽  
Y. Aoyagi
Keyword(s):  
Sex Ratio ◽  
Animal Health ◽  
Transrectal Ultrasonography ◽  
Control Group ◽  
Chi Square ◽  
Female Ratio ◽  
Significant Difference ◽  
Chi Square Test ◽  
Rectal Palpation ◽  
And Control

Skewing the sex ratio of offspring towards males or females is very important for the livestock industry. Many factors, such as maternal stress, have been suggested to affect the sex ratio (Pratt NC et al. 1989 J. Reprod. Fertil. 87, 763–769). In a recent study (Ideta A et al. 2007 J. Reprod. Dev. doi:10.1262/JRD.19035), the proportion of female embryos recovered from superovulated heifers in which ovulation patterns were observed by repeated transrectal ultrasonography tended to be higher than the expected ratio of 50:50 (66.7%, 26/39). To investigate this phenomenon, we repeated the experiment using a larger number of Holstein heifers. The superovulatory treatment began in the midluteal phase of the estrous cycle (days 8 to 10) and consisted of eight decreasing doses of FSH i.m. (total of 28 Armour units, Antrin R-10, Kawasaki-Mitaka, Kanagawa, Japan) for 4 days with treatment twice daily. Doses of 5 mL and 3 mL of a PGF2α analogue (Veterinary Pronalgon F Injection containing 5 mg mL–1 Dinoprost, Pfizer Animal Health, Tokyo, Japan) were administered i.m. to the animals along with the seventh and eighth FSH treatment, respectively. The heifers were divided into two groups. One group, the rectal palpation (RP) group (n = 9), received transrectal ultrasonography with rectal palpation at 4-h intervals from 36 to 76 h after the first PGF2α treatment. The other group, the Control group (n = 8) received no treatment. The heifers were artificially inseminated at 56 and 72 h after the first PGF2α treatment using frozen–thawed semen from one bull. Seven-day embryos were recovered nonsurgically. Grade 1 to 3 embryos (IETS classification) were selected for this study. Male and female embryos were separated using the loop-mediated isothermal amplification procedure (Hirayama H et al. 2004 Theriogenology 62, 887–896). Data were analyzed using ANOVA and chi-square test. The mean number of recovered ova and embryos was 15.7 � 3.8 (RP) and 14.4 � 2.2 (Control). There was no significant difference in the percentages of unfertilized ova (RP; 14.9 %, 21/141 and Control; 11.3% 13/115, P > 0.05), grade 1 embryos (RP; 51.1%, 72/141 and Control; 54.8%, 63/115, P > 0.05) and grade 1 to 3 embryos (RP; 65.2%, 92/141 and Control; 69.6%, 80/115, P > 0.05) between the two groups. The proportion of female grade 1 embryos in the RP group (66.7%, 48/72) was significantly higher than the expected ratio of 50:50 (P < 0.01). The female ratio of grade 1 embryos in the Control group was 50.8% (32/63). Furthermore, the proportion of female grade 1 to 3 embryos in the RP groups (66.3%, 61/92) was significantly higher than the expected ratio of 50:50 (P < 0.005). The female ratio of grade 1 to 3 embryos in the Control group was 51.3% (41/80). Results indicate that frequent ultrasound examinations and rectal palpations following superovulatory treatment may skew the sex ratio of embryos towards females in Holstein heifers.

318 EFFECT OF SPERM PRETREATMENT WITH ISOBUTYLMETHYLXANTHINE AND METHYL-β-CYCLODEXTRIN ON THE EFFICIENCY OF BOVINE INTRACYTOPLASMIC SPERM INJECTION

2015 ◽  
Vol 27 (1) ◽  
pp. 248
Author(s):  
L. M. Aguila ◽  
M. E. Arias ◽  
R. S. Sanchez ◽  
T. C. Vargas ◽  
F. A. Zambrano ◽  
...  
Keyword(s):  
Intracytoplasmic Sperm Injection ◽  
Defined Medium ◽  
Fertilization Rate ◽  
Vehicle Group ◽  
Control Group ◽  
Sperm Capacitation ◽  
Chi Square ◽  
Developmental Potential ◽  
Pronuclear Formation

The efficiency of intracytoplasmic sperm injection (ICSI) in bovines is lower than in other species. We propose that in vitro sperm capacitation could optimize the ICSI in cattle. The aim was to evaluate the effects of isobutylmethylxanthine (IBMX) and methyl-β-cyclodextrin (MβCD) on the sperm capacitation and in vitro development of embryos generated by ICSI. Frozen-thawed spermatozoa (3–5 × 106 cells mL–1) were pre-incubated for 2 h at 38.5°C, 5% CO2 in defined medium (Sp-TLP/PVA) supplemented with MβCD (1 mM) or IBMX (0.4 mM) (capacitating conditions). The untreated control group (UTG; not supplemented) and vehicle group (VG) were incubated for 2 h. The non-capacitating control group (NCG) was not supplemented (neither vehicle nor IBMX or MβCD) and not incubated. The sperm viability and capacitation {intracellular calcium [Ca2+]i, plasma membrane fluidity (PMF), and acrosomal reaction} were evaluated by flow cytometry (n = 3 biological replicates). For the ICSI procedure, only motile spermatozoa were selected. After ICSI, oocytes were activated with ionomycin + cycloheximide. Culture was performed at 38.5°C, 5% CO2, 5% O2, 90% N2, saturation humidity in KSOM base medium. Data were analysed by ANOVA and Scheffe's test. Pronuclear formation was evaluated by a chi-square test with Bonferroni's correction. Significance was set at P < 0.05. Pretreated spermatozoa showed lower (P < 0.05) viability (49 and 67% for IBMX and MβCD, respectively) compared with the NCG (89%), UTG (80%), and VG (78%). The [Ca2+]I analysed by median fluorescence intensity (MFI) was lower (P < 0.05) in NCG (117 MFI) with respect to UTG (127 MFI), VG (124 MFI), IBMX (126 MFI), and MβCD (131 MFI). The PMF increased (P < 0.05) with IBMX (115 MFI) and MβCD (106 MFI) compared with NCG (70 MFI), UTG (89 MFI), and VG (65 MFI). Acrosome reaction improved with capacitating treatments with respect to both control groups (16, 23, 8, 4, and 3% for IBMX, MβCD, UTG, VG, and NCG, respectively). Analysis of capacitating v. non-capacitating conditions on ICSI efficiency revealed that the fertilization rate, assessed by pronuclear formation, was higher (P < 0.05) in ICSI-MβCD (76%; n = 46) compared with ICSI-IBMX (55%; n = 53) and ICSI-NCG (50%; n = 44). Nevertheless, there were no differences among groups in cleavage (Day 3): 85, 86, and 84% and blastocyst rates (Day 8): 19, 25, and 18% for ICSI-IBMX (n = 8), ICSI-MβCD (n = 7), and ICSI-NCG (n = 7), respectively. The parthenogenetic and sham injection groups yielded a lower rate of cleavage (73 and 53%, respectively) and blastocyst (13% and 10%, respectively). The results demonstrated an improvement of the fertilization rate of bovine embryos generated by ICSI using sperm capacitated by MβCD pretreatment. However, more studies are necessary to improve in vitro developmental potential of these embryos to the blastocyst stage.Frigorífico Temuco and funding support from FONDECYT 1120241 CONICYT-Chile are gratefully acknowledged.

362 SEX RATIO OF IN VITRO-PRODUCED BLASTOCYSTS OBTAINED USING FROZEN SEMEN FROM A CLONED BULL OR ITS ORIGINAL CELL DONOR

2007 ◽  
Vol 19 (1) ◽  
pp. 296
Author(s):  
Y. Heyman ◽  
B. LeGuienne ◽  
C. Audouard ◽  
F. Charreaux ◽  
P. Humblot ◽  
...  
Keyword(s):  
Sex Ratio ◽  
Vero Cells ◽  
Chi Square ◽  
Cell Donor ◽  
Frozen Semen ◽  
Chi Square Test ◽  
Original Cell ◽  
Vitro Development ◽  
Original Donor

Bulls obtained by somatic cell nuclear transfer (SCNT) have proved to develop normally, produce sperm, and be fertile (Shiga et al. 2005 Theriogenology 64, 334–343; Tecirlioglu et al. 2006 Theriogenology 65, 1783–1799). However, due to epigenetic variability encountered with clones, it cannot be excluded that major alterations may affect the X chromosome and especially the dosage of X inactivation in female embryos. This could result in a deviation of the sex ratio in offspring. To test this hypothesis, the sperm from a cloned bull was used in IVF to assess its ability to induce a different proportion of male and female embryos when compared to results obtained with the sperm of the original donor before cloning. In the present experiment, semen was collected twice weekly during 3 months from a 3-year-old cloned bull of the Charolais breed (H-2) and frozen-stored. Samples from 3 different ejaculates were used in comparison with control frozen straws from the original bull (H-0), prepared at the same age, to produce in vitro embryos. A total of 6 replicate IVF experiments were performed on the same batches of in vitro-matured oocytes (n = 654) using the standard swim-up and IVF technique in the laboratory. Twenty hours after insemination, presumptive zygotes were vortexed and cultured for 7 days in microdrops of B2 medium with Vero cells. Fertilization, cleavage, and blastocyst formation was assessed, and by Day 7, all of the blastocysts were individually frozen after grading. Two groups of 40 representative blastocysts derived from each bull (clone or donor) were used to determine the sex ratio using the sexing kit developed by UNCEIA R&amp;D (Maisons-Alfort, France). Percentages between groups were compared by chi-square test. Semen from the cloned bull resulted in significantly lower fertilization and cleavage rates than that from the original donor (82.7% and 70% vs. 94.6% and 91%, respectively; P &lt; 0.01), but further in vitro development was not different, as the proportions of blastocysts/cleaved were, respectively, 31.5% (74/235) vs. 38.7% (112/290). Sex determination was achieved in 79/80 of the in vitro-produced embryos and indicated that 55% of the blastocysts derived from the clone were male (22/40) and 45% female (18/40). This was not different from the proportion of male (61.5%, 24/39) and female (38.5%, 15/39) embryos in the group derived from semen of the original donor bull. In conclusion, these preliminary results indicate that the semen from this cloned Charolais bull has the same potential for in vitro embryo production as its original cell donor, and there is no evidence that SCNT could induce any deviation of the sex ratio. This observation needs to be confirmed with other sets of cloned bulls. (Shiga et al. 2005 Theriogenology 64, 334–343. Tecirlioglu et al. 2006 Theriogenology 65, 1783–1799.)

scholarly journals 46 Effect of the addition of α-tocopherol to in vitro maturation media of bovine oocytes

2020 ◽  
Vol 98 (Supplement_2) ◽  
pp. 2-3
Author(s):  
Theisy P Acosta Pérez
Keyword(s):  
In Vitro Maturation ◽  
Cumulus Cells ◽  
Control Group ◽  
Chi Square ◽  
Cumulus Expansion ◽  
Minimum Concentration ◽  
Maturation Rate ◽  
Chi Square Test ◽  
System Data

Abstract α-tocopherol is known to be a powerful antioxidant, in this regard, it was added to bovine oocyte in vitro maturation media to evaluate its effect on oocyte maturation. Oocytes (n = 624) aspirated from ovaries of slaughtered cows were classified by quality and divided in four categories according to cytoplasm appearance and cumulus cells layers. Oocytes were washed in TCM-199 supplemented with fetal bovine serum (FBS) and FSH, then distributed in maturation media (TCM-199 supplemented with FBS, FSH and gentamicin). Three experimental groups of α-tocopherol (50, 100 and 200 mM) and a control group without α-tocopherol were used. Maturation was carried 22 h at 38.5°C in a 5% CO2 atmosphere. Oocytes were examined to determine cumulus expansion as categorical data (expansion or no expansion), as well as cumulus expansion Index (CEI). For CEI determination oocytes were graded 0 to 4, being 0 those with null expansion and 4 those with a noticeable cell expansion, then the number of oocytes were multiplied by the grade given and a sum of the totals was obtained, the new total was divided by the total of oocytes in the group and the result obtained corresponded to the CEI of the group. Results were analyzed with Chi Square test (for maturation rates) and an ANOVA (for the CEI) using the SAS system, data are presented as mean ± standard error. There was no statistical difference between control and α-tocopherol groups (P &gt;0.05). Numerically, the control group showed a higher maturation rate (100%) and obtained a higher CEI (2.44±0.20), followed by the 50 mM group (98.16%; 2.39±0.13), the groups 200 mM (97.40%; 2.00±0.14) and 100 mM (96.25%; 2.06±0.24) were the lowest. The addition of the minimum concentration (50 mM) of α-tocopherol to the maturation media could improve maturation rates without exposing oocytes to toxic effects.

Effect of vitrification technique and assisted hatching on rabbit embryo developmental rate

Zygote ◽  
2009 ◽  
Vol 17 (1) ◽  
pp. 57-61 ◽  
Author(s):  
M. Popelková ◽  
Z. Turanová ◽  
L. Koprdová ◽  
A. Ostró ◽  
S. Toporcerová ◽  
...  
Keyword(s):  
Cell Number ◽  
Total Cell ◽  
Control Group ◽  
Assisted Hatching ◽  
Hatching Rate ◽  
Total Cell Number ◽  
Developmental Potential ◽  
Significant Difference ◽  
Rabbit Embryos

SummaryThe aim of the study was to determine the efficiency of two vitrification techniques followed by two assisted hatching (AH) techniques based on post-thaw developmental capacity of precompacted rabbit embryos and their ability to leave the zona pellucida (hatching) during in vitro culture. The total cell number and embryo diameter as additional markers of embryo quality after warming were evaluated. In vivo fertilized, in vitro cultured 8–12-cell rabbit embryos obtained from superovulated rabbit does were cryopreserved by two-step vitrification method using ethylene glycol (EG) as cryoprotectant or by one-step vitrification method with EG and Ficoll (EG+Ficoll). Thawed embryos were subjected to enzymatic or mechanical AH. Vitrified EG group showed significantly lower (P < 0.05) blastocyst rate (22.5%) and hatching rate (15%) than those vitrified with EG + Ficoll (63 and 63% resp.) and that of control (97 and 97% respectively). Significantly lower values of total cell number (P < 0.05) as well as embryo diameter (P < 0.01) in EG group compared with EG + Ficoll and control group were recorded. No significant difference was found in developmental potential of warmed embryos treated by either mechanical or enzymatic AH. The present study demonstrates that the EG + Ficoll vitrification protocol provides superior embryo survival rates over the EG vitrification protocol for 8–12-cell stage precompacted rabbit embryos. No positive effect of either mechanical or enzymatic AH on the post-thaw viability and quality of rabbit embryos in vitro was observed.

scholarly journals In vitro activity of zinc oxide-eugenol and glass ionomer cements on Candida albicans

2005 ◽  
Vol 19 (2) ◽  
pp. 134-138 ◽  
Author(s):  
Anna Carolina Aguiar Cassanho ◽  
Aletéia Massula Fernandes ◽  
Luciane Dias de Oliveira ◽  
Claudio Antonio Talge Carvalho ◽  
Antonio Olavo Cardoso Jorge ◽  
...  
Keyword(s):  
Zinc Oxide ◽  
Candida Albicans ◽  
Control Group ◽  
Glass Ionomer ◽  
Experimental Conditions ◽  
Mean Values ◽  
Zinc Oxide Eugenol ◽  
Colony Forming Units ◽  
And Control

The aim of this study was to evaluate in vitro the antimicrobial activity of glass ionomer (GIC) and zinc oxide-eugenol (ZOE) cements against Candida albicans. Standardized GIC and ZOE specimens were maintained in contact with C. albicans suspension (1 <FONT FACE=Symbol>´</FONT> 10(6) cells/ml) at 37°C for 24 h, 48 h or 7 days. A control group without any testing cement was included. After the incubation period, aliquots of 0.1 ml were plated on Sabouraud's agar, and then the number of colonies was counted. The results were expressed as values of logarithms of colony-forming units per milliliter (log CFU/mL) and were analyzed statistically by Kruskal-Wallis ANOVA. After 48 h of incubation, the ZOE group presented no growth of C. albicans. GIC and control groups presented similar mean values at all tested periods. According to the results obtained, it could be concluded that, under the experimental conditions, ZOE cement was more effective in vitro against C. albicans than GIC.

PSX-B-11 Comparing effects of FGF2 and IGF1 on the development competence of parthenogenetic activated bovine oocytes maturing in vitro

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 327-328
Author(s):  
Galina Singina
Keyword(s):  
Growth Factor ◽  
Cell Number ◽  
Blastocyst Stage ◽  
Total Cell ◽  
Control Group ◽  
Total Cell Number ◽  
Developmental Potential ◽  
Maturation Medium ◽  
Bovine Oocytes

Abstract The oocyte quality acquired during in vitro maturation (IVM) are the main limitative factors affecting the embryo production. The aim of the present research was to study effects of fibroblast growth factor 2 (FGF2) and insulin-like growth factor 1 (IGF1) during IVM of bovine oocytes on their developmental potential after parthenogenetic activation. Bovine cumulus-oocyte complexes (COC; n = 1176) were cultured for 22h in either standard maturation medium (TCM-199 supplemented with 10% fetal calf serum (FCS), 0.2 mM sodium pyruvate, 10 μg/ml FSH and 10 μg/ml LH; Control) or maturation medium supplemented with different concentrations (5–160 ng/ml) of FGF2 and IGF1. After IVM, matured oocytes activated by sequential treatment with ionomycin followed by DMAP and cyclohexamide and then cultured up to the blastocyst stage. The obtained blastocysts were fixed, and the total cell number and the level of apoptosis were determined using DAPI and TUNEL staining. The data from 4 replicates (77–91 oocytes per treatment) were analyzed by ANOVA. Cleavage rates of activated oocytes did not differ between groups and ranged from 63.7 to 68.1%. The addition of 10, 20 and 40 ng/ml of FGF2 to the IVM medium led to an increase in the yield of blastocysts [from 19.6±1.8% (Control) to 35.2±3.4, 29.8±1.9 and 31.1±2.1%, respectively (P&lt;0.05)] and in the total cell number in embryos that developed to the blastocyst stage (P&lt;0.05). Meanwhile, the blastocyst yield and the total cell number in blastocysts in the IGF1-treated groups were similar to that in the control group. No effects of both growth factors on the proportion of apoptotic nuclei in blastocysts (5.3–7.1%) were observed. Thus, FGF2 (but not IGF1) are able to maintain competence for parthenogenetic development of bovine COC during their maturation invitro. Supported by RFBR (18-29-07089) and the Ministry of Science and Higher Education of Russia.

Improvement of the developmental competence of canine oocyte using caffeine supplementation during IVM at different maturation time

Zygote ◽  
2018 ◽  
Vol 26 (2) ◽  
pp. 162-167 ◽  
Author(s):  
Mohamed Fathi ◽  
A. Salama ◽  
Magdy R. Badr
Keyword(s):  
Developmental Competence ◽  
Control Group ◽  
Free Medium ◽  
Maturation Time ◽  
Fluid Medium ◽  
Developmental Potential ◽  
Nuclear Maturation ◽  
Maturation Medium ◽  
Oviductal Fluid

SummaryThe aim of the current study was to investigate the effect of caffeine supplementation during in vitro maturation (IVM) for different maturation times on the developmental potential of canine oocytes recovered from ovariohysterectomized bitches. The recovered cumulus–oocytes complexes were in vitro matured for 72 h. Here, 10 mM caffeine was added to the maturation medium for different incubation times (caffeine from 0–72 h maturation, caffeine for the first 24 h of maturation only, caffeine addition from 24 to 48 h maturation time, caffeine addition from 48 to 72 h maturation or in caffeine-free medium, control group). The matured oocytes were in vitro fertilized using frozen–thawed spermatozoa. The presumptive zygotes were in vitro cultured in synthetic oviductal fluid medium for 5 days. The results showed that both maturation and fertilization rates were significantly higher (P ˂ 0.05) using caffeine-treated medium for the first 24 h of maturation compared with the control and other two groups of caffeine treatment (from 24 to 48 h and from 48 to 72 h), whereas use of caffeine-treated medium for a 0–72 h incubation time did not affect these rates (P > 0.05). Interestingly, the matured oocytes in caffeine-supplemented medium for the first 24 h or from 0–72 h showed a significant (P ˂ 0.05) increase in the total number of cleaved embryos compared with the control group. In conclusion, supplementation of the maturation medium with 10 mM caffeine for the first 24 h of maturation or during the whole maturation time (0–72 h) improved nuclear maturation and subsequent embryo development preimplantation following in vitro fertilization.

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