Fertilization, sperm transport, and early embryonic loss in ewes grazed on cultivars of subterranean clover. (T. subterraneum)

1970 ◽  
Vol 21 (1) ◽  
pp. 107 ◽  
Author(s):  
cC Kaltenbach ◽  
HL Davies

The causes of infertility due to phyto-oestrogens were investigated in ewes which had grazed Dwalganup, Yarloop, Geraldton, Woogenellup, and Mount Barker cultivars of subterranean clover (T. subterraneum L.) for 5 years. At laparotomy, sperm transport was investigated by making counts of spermatozoa in the oviduct 22-26 hr post coitum; fertilization rates were checked 72 hr post coitum and embryonic survival at 27-31 days. A larger proportion of ewes on high oestrogen cultivars Dwalganup, Yarloop, and Geraldton) had low oviducal spermatozoa counts and the fertilization rate was significantly lower (P < 0.05) than in ewes grazed on low oestrogen cultivars (Woogenellup and Mount Barker) (44% v. 63 %). There was no statistically significant difference in embryonic survival between these groups. Merino ewes had fewer oviducal spermatozoa and a lower (P < 0.05) fertilization rate than Border Leicester x Merino ewes. Impaired sperm transport is considered to be the reason for reproductive failure in the ewes that had grazed the high oestrogen cultivars of subterranean clover.

1968 ◽  
Vol 19 (6) ◽  
pp. 1059 ◽  
Author(s):  
HE Fels ◽  
HG Neil

Six-year-old ewes, the survivors of two experimental groups that had grazed from May to October through the 5 preceding years on mixed pastures containing an oestrogenic variety (Dwalganup) of subterranean clover ("clover" group) and oat crops ("oats" group), were mated 6 months after their last exposure to oestrogenic pasture and allocated for slaughter to observe the fertilization rate of eggs or the numbers of surviving embryos. In late April the clover and oats groups did not differ significantly in the proportion of eggs fertilized (13/15 eggs v. 11/11) but in May there was a highly significant difference (4/12 v. 14/17). The difference between late April and May in the clover group was significant and was associated with changes in the ratio of oestrous ewes to rams. The ratio of live embryos to eggs shed was much higher in the oats than in the clover group (56173 v. 8/39). After estimating how many of these eggs had been fertilized it was concluded that more fertilized eggs developed successfully in the oats than in the clover group (c. 56/65 v. 8/17). The data suggested that more "clover" embryos died during days 13–19 of pregnancy. Significantly more clover group ewes had obvious reproductive tract infections. These ewes did not show oestrus regularly though they may have ovulated more or less regularly. Other clover group ewes showed oestrus as regularly as oats group ewes. Clover group ewes had many more macroscopic endometrial cysts in their uteri and cervices, but within groups, numbers of cysts were not significantly associated with previous or current reproductive performance.


2001 ◽  
Vol 26 (1) ◽  
pp. 161-174 ◽  
Author(s):  
J.C. Dalton ◽  
S. Nadir ◽  
J. Bame ◽  
M. Noftsinger ◽  
R.G. Saacke

AbstractTo further identify factors which influence pregnancy rates, three experiments were conducted to determine the effect of insemination time on sperm transport, fertilization rate, and embryo quality. All cows were continuously monitored for behavioural oestrus by HeatWatch®, and received AI at heat onset (0 h after the first standing event), 12 h after onset, or received natural service at 0 hfrom one of three bulls (Exp. 1). In Exp. 2, cows received AI at 0 h, 12 h, or 24 h after the first standing event. On d 6 after insemination 115 embryos(ova) (Exp. 1) and 117 embryos(ova) (Exp. 2) were recovered from single-ovulating cows. For Exp. 1, median accessory sperm values were: 1 (0 h), 10 (12 h), 27 (natural service O h) (P < 0.05). For Exp. 2, median accessory sperm values were: 1 (0 h), 2 (12 h), 4 (24 h) (P < 0.05). Fertilization rates were: 67% (0 h), 79% (12 h), 98% (natural service O h) (P < 0.05)(Exp. 1); and did not differ in Exp. 2. Embryo quality was not different in Exp. 1. In Exp. 2, percentages of excellent and good fair and poor, and degenerate embryos were: 77, 15, 8 (0 h), 52, 38, 10 (12 h), 47, 19, 34 (24 h) (P < 0.05). In Exp. 3, 30 cows were superovulated and were inseminated once at either 0 h, 12 h, or 24 h after the first standing event. On d 6 after insemination, 529 embryos(ova) were recovered. Fertilization rates were: 29% (0 h); 60% (12 h); 81% (24 h)(P < 0.01). Percentages of embryos with accessory sperm were: 5 (0 h); 8 (12 h); and 41(24 h) (P < 0.01). Embryo quality was not affected by time of AI. We conclude that the time of insemination affects: 1) sperm transport as measured by median accessory sperm number (Exp. 1 and 2) and the percentage of embryos with accessory sperm (Exp. 3); 2) fertilization rate (Exp. 1 and 3); and embryo quality (Exp. 2).


1971 ◽  
Vol 11 (52) ◽  
pp. 478
Author(s):  
JR Giles

A series of studies was conducted in 1968 and 1969 to compare fertilization rates, return rate, and sperm distribution in the genital tract in Bungaree and Peppin Merino ewes. Differences in lambing rates of the two ewe strains were demonstrated, but there was no major difference in fertilization rate in either year. A significantly higher proportion of Bungaree ewes had less than 1000 sperm in the fallopian tubes 24 hours after service, but there were no significant differences in any other character examined. Contrary to an earlier report, fertilization failure is unlikely to have been a major cause of a difference in the fertility of the two strains in the two years, and embryonic loss is likely to have made a significant contribution to the difference in their fertility.


2012 ◽  
Vol 24 (1) ◽  
pp. 192 ◽  
Author(s):  
A. R. Moawad ◽  
G. M. Darwish ◽  
M. R. Badr ◽  
A. B. El-Wishy

Various techniques such as AI and ET have been reported to improve reproductive efficiency and genetic potential in camelids. In vitro fertilization and the development of IVP embryos are considered an alternative for genetic improvement in this species. This study investigated the effects of different sperm cell concentrations (1, 2, 3 and 4 × 106 sperm mL–1), different capacitating materials (5 mM caffeine, 10 μg mL–1 of heparin, 10 mg mL–1 of theophylline, 1 mM calcium ionophore A23178 and 10 μg of heparin + 5 mM caffeine), post-slaughter epididymal flushing time and fertilization media supplements (Fert-TALP + 6 mg mL–1 of BSA and Fert-TALP + 3 mg mL–1 of polyvinylpyrrolidone ) on fertilization rates and subsequent development of dromedary camel oocytes. Cumulus–oocyte complexes obtained at slaughter were matured in vitro in TCM-199 for 36 h at 39°C in a humidified atmosphere of 5% CO2. For IVF, spermatozoa were collected from epididymides of slaughtered male camels at 1 to 2 h post-slaughter or after 24 h of epididymal storage at 4°C. The spermatozoa were then prepared for IVF by the swim-up technique. Following sperm capacitation, oocytes and spermatozoa were co-incubated for 18 h. Oocytes were then stained using aceto-orcein for evaluation of fertilization events. Presumptive zygotes were cultured in vitro in TCM-199 medium supplemented with 5% FCS for 9 days at 39°C in a humidified atmosphere of 5% O2, 5% CO2 and 90% N2. At least 3 replicates were performed for each experimental group. Data were analysed by chi-square test. Fertilization rates were 55.5, 62.5, 62.7 and 47.2% in oocytes inseminated with 1, 2, 3, or 4 × 106 sperm mL–1, respectively. Normal fertilization rate (oocytes with 2 pronuclei) was higher (P =  0.06) in oocytes inseminated with 2 × 106 sperm mL–1 (29.7%) than in those inseminated by 4 × 106 sperm mL–1 (11.1%). Treatment of epididymal spermatozoa with 5 mM caffeine significantly increased (P ≤ 0.05) fertilization rate (61.9%) compared with calcium ionophore A23178 (32.4%). These values were not significantly different from other groups (38.5, 54.1 and 50.0% in heparin, theophylline and heparin + caffeine, respectively). Normal fertilization was highest (25.4%) in oocytes inseminated with caffeine-treated spermatozoa. Insemination of oocytes in Fert-TALP medium containing BSA resulted in a higher fertilization rate (21.4%) compared with oocytes in polyvinylpyrrolidone-supplemented medium (5.7%; P =  0.06). Storage of camel epididymides at 4°C for 24 h did not affect fertilization rates. Cleavage rate (48 h post-insemination) was higher in oocytes fertilized with caffeine-treated spermatozoa than in oocytes in the theophylline group (26.8 vs 10.5%; P =  0.08). No significant difference was observed in the frequency of blastocyst development (5 days post-insemination) between the 2 groups (5.4 vs 2.6%); based on the number of cleaved oocytes, the same proportions of blastocyst embryos were reported (20.0 and 25.0%). Taken together, these results suggest that dromedary camel oocytes can be matured, fertilized and subsequently developed in vitro with high developmental potential. Epididymal spermatozoa at a concentration of 2 × 106 sperm mL–1 prepared in a medium containing caffeine as a capacitating agent can be used effectively in IVF of camel oocytes.


2013 ◽  
Vol 25 (1) ◽  
pp. 159
Author(s):  
F. Becker ◽  
W. Kanitz ◽  
G. Nurnberg ◽  
D. Rath

Different factors determine the effectiveness of the use of sires in AI. Most important factors are the number of inseminated spermatozoa, the quality of spermatozoa, and the time of insemination. Especially in superovulated animals, the insemination scheme plays in important role to cover the whole ovulation period. The influence of 3 different dosages of spermatozoa (15 × 106, 5 × 106, and 1 × 106) on fertilization rate was examined in experiment A. In experiment B, one dosage of female and male spermatozoa of 3 different bulls was used for timed AI in 31 heifers. Timed AI in normal-cycling cattle [13 h after gonadotropin-releasing hormone (GnRH) application] with detected corpus luteum (Days 8 to 13 of cycle) was carried out after induction of luteolysis and induction of ovulation [GnRH application 60 h after prostaglandin F2α (PGF2) application]. Embryos and oocytes were flushed from the oviduct of 116 hemicastrated or slaughtered heifers on Day 4 after insemination. The ovulation rate in heifers was 95.4%. Eighty percent of the oocytes or embryos were recovered. The influence of the factors sire, ejaculate, and dosage were tested by GLM analyses of SAS® (SAS Institute Inc., Cary, NC, USA). There was no significant difference in the fertilization rate (93.3, 96.2, and 78.8%) and in the proportion of normally developed embryos (84.6, 80.7, and 75.8%) between groups. Significant differences were found in the mean number of accessory sperms/embryo and in the proportion of embryos with >10 accessory sperms/embryo or without accessory sperms; however, the proportion of intact embryos was similar. Using sexed semen in experiment B, similar results were obtained after flushing of the oviducts on Day 4 after insemination of hemicastrated or slaughtered animals. In total, an ovulation rate of 91.7%, a recovery rate of 70%, and a fertilization rate of 86.8% were obtained. There were no differences between female- and male-sorted spermatozoa and the control group. In experiment C, altogether 13 heifers were treated 8 times with FSH for 4 days starting between Day 8 to 12 of estrous cycle. Prostaglandin F2α was given 48 and 60 h after the first FSH injection. Insemination with sexed semen (n = 5 heifers) and with unsorted semen (n = 8; 15 × 106 and 1 × 106) was done at 55 and 71 h after induction of luteolysis. Flushing of the uterus was performed on Day 7. Using the time-oriented insemination after superovulation of animals, fertilization rates varied between 65 and 85%. There was no difference between groups regarding the number of transferable embryos (5.5, 4.9, and 4.8). The results demonstrate that the application of an approved insemination schedule may accomplish high fertilization rates after insemination with sexed or reduced dosages of spermatozoa in normal-cycling as well as superovulated cattle.


2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
A Schallmoser ◽  
J Verguts ◽  
J P Allam ◽  
N Sänger

Abstract Study question The main objective of this study was to examine the influence of mild α -chymotrypsin treatment of highly viscous semen samples on the fertilisation rate after artificial reproductive treatment (ART). Summary answer The use of mild α -chymotrypsin treatment of semen samples in case of hyperviscosity does not appear to impact negatively on the fertilization rates after ART. What is known already Highly viscous semen reduces sperm motility significantly and can contribute to infertility. When processing semen samples, few techniques exist to induce liquefaction in case of seminal hyperviscosity such as different washing steps and mechanical treatment. The use of α -chymotrypsin seems controversial due to possible negative effects on fertilisation rates after IVF. Study design, size, duration All patients were recruited at the Department of Gynecological Endocrinology and Reproductive Medicine, University Hospital of Bonn, Germany from July first 2018 till June first 2019. Anonymized data on age, number of previous pregnancies and deliveries were retrospectively collected. The study group consisted of a cohort of 52 couples meeting the inclusion criteria of highly viscous semen and were compared to a cohort control group of 88 couples. Participants/materials, setting, methods: The fertilization rate of 52 ART cycles was examined following IVF using a low dose of α -chymotrypsin to induce liquefaction of highly viscous semen and was compared to a control group of 88 ART cycles. Data was analyzed using SPSS version 25. A Mann Whitney U test was used to compare continuous parameters between groups. Main results and the role of chance The study group consisted of a cohort of 52 couples meeting the inclusion criteria of highly viscous semen and were compared to a cohort control group of 88 couples. The Fertilization rate between the two groups was not significant (p &lt; 0.146, Mann-Whitney U test), with a rate of 57.95 in the study group compared to 57.53 in the control group. Our analysis showed no significant differences in male and female age, male abstinence period, semen volume, sperm concentration, total sperm count, and total progressive sperm count between the two groups. We observed a significant difference [p = 0.025] in the progressive motility and a borderline significance in the total progressive sperm count [p = 0.052] between α -chymotrypsin treated samples and the non α -chymotrypsin treated samples. Analyzing the clinical and biochemical pregnancy parameters of the study group (n = 41) and the control group (n = 66) per fresh embryo transfer procedures (n = 107) we found no significant differences. Freeze all cycles were excluded from the study. No significant differences concerning previous pregnancies and childbirth were detected. Limitations, reasons for caution The study is a pilot study as the majority of the studies using α -chymotrypsin was conducted up to three decades ago, comparison of data must be interpreted in the light of the fact that since then, IVF has seen a rapid evolution of technology and culture techniques. Wider implications of the findings The use of mild α -chymotrypsin treatment of semen samples in case of hyperviscosity does not appear to impact negatively on the fertilization rates after ART and could be regarded as an additional method to induce liquefaction of highly viscous semen samples in IVF... Trial registration number not applicable


2004 ◽  
Vol 94 (5) ◽  
pp. 433-439 ◽  
Author(s):  
S.N. Johnson ◽  
P.J. Gregory ◽  
P.J. Murray ◽  
X Zhang ◽  
I.M. Young

AbstractThis study investigated the ability of neonatal larvae of the root-feeding weevil, Sitona lepidus Gyllenhal, to locate white clover Trifolium repens L. (Fabaceae) roots growing in soil and to distinguish them from the roots of other species of clover and a co-occurring grass species. Choice experiments used a combination of invasive techniques and the novel technique of high resolution X-ray microtomography to non-invasively track larval movement in the soil towards plant roots. Burrowing distances towards roots of different plant species were also examined. Newly hatched S. lepidus recognized T. repens roots and moved preferentially towards them when given a choice of roots of subterranean clover, Trifolium subterraneum L. (Fabaceae), strawberry clover Trifolium fragiferum L. (Fabaceae), or perennial ryegrass Lolium perenneL. (Poaceae). Larvae recognized T. repens roots, whether released in groups of five or singly, when released 25 mm (meso-scale recognition) or 60 mm (macro-scale recognition) away from plant roots. There was no statistically significant difference in movement rates of larvae.


2005 ◽  
Vol 17 (2) ◽  
pp. 159 ◽  
Author(s):  
E.A. Martinez ◽  
J.M. Vazquez ◽  
I. Parrilla ◽  
C. Cuello ◽  
M.A. Gil ◽  
...  

A new procedure for nonsurgical deep intrauterine insemination (DUI) in non-sedated sows has recently been reported (Martinez et al. 2002 Reproduction 123, 163–170). In comparison to traditional artificial insemination (AI), using this procedure, a 20-fold reduction in the number of spermatozoa inseminated can be used without a decrease in fertility when hormonally treated post-weaning estrous sows are used. The aim of the present study was to evaluate the effectiveness of DUI under field conditions. In Experiment 1, crossbred sows (2–6 parity) were weaned at 20.75 ± 0.06 days. Estrous detection was performed once per day, beginning 3 days after weaning. Sows with a weaning to estrus interval of 4–5 days were selected to be inseminated. A total of 190 sows were inseminated at 12, 24, and 36 h after onset of estrus using one of the following two regimes: (1) DUI with 150 × 106 fresh spermatozoa in 5 mL of BTS (n = 95) and (2) Traditional AI with 3 × 109 fresh spermatozoa in 100 mL of BTS (n = 95) prepared from the same semen samples used for the DUI group. Farrowing rates (FR) and litter sizes (LTS; mean ± SEM) from both groups were compared using chi-squared test and ANOVA, respectively. There was no significant difference in the FR between groups (83.2 and 86.3% for DUI and AI groups, respectively). However, a decrease (P < 0.001) in the LTS was observed in sows inseminated by the DUI procedure (9.8 ± 0.29 and 10.9 ± 0.17, respectively). In Experiment 2, seventy one natural post-weaning estrus sows were used. Fifty-five sows were DUI inseminated three times with 150 (n = 17), 300 (n = 19), or 600 (n = 19) × 106 spermatozoa in 5, 10, or 20 mL of BTS, respectively. The remaining sows (n = 16) were traditionally inseminated. On Day 6 after estrus, sows were subjected to laparotomy and the tips of both uterine horns were flushed in order to evaluate pregnancy rate (PR: percentage of sows with at least 4 viable embryos) and fertilization rate (ratio of viable embryos to the total number of embryos and oocytes). PR was similar in all the groups, ranging from 84.2% (DUI 300 × 106 spermatozoa group) to 94.7% (DUI 600 × 106 spermatozoa group). Fertilization rate and the percentage of bilateral fertilization after DUI with 600 × 106 spermatozoa did not differ from those of the AI group (97.8 and 100% vs. 98.4 and 100%, respectively), but a significant decrease in both parameters (P < 0.05; chi-square test) was observed in sows inseminated with 300 (94.3 and 87.5%) or 150 (84.4 and 66.7%) × 106 spermatozoa. In conclusion, DUI with 150 × 106 spermatozoa offers similar FR but a lower LTS in sows with natural estrus in comparison with those parameters obtained when traditional AI is used. The lower litter size could be related to the low percentage of bilateral fertilization observed in that group. This work was supported by CDTI 020003.


2021 ◽  
Author(s):  
Jian Xu ◽  
Li Yang ◽  
Zhi-Heng Chen ◽  
Min-Na Yin ◽  
Juan Chen ◽  
...  

Abstract Objective: To investigate whether the euploidy rate of blastocysts derived from smooth endoplasmic reticulum (SERa) positive cycles and oocytes are impacted.Design: Retrospective cohort study.Setting: A tertiary hospital-based reproductive medicine center.Patient(s): A total of 601 preimplantation genetic testing (PGT) cycles with obtained oocytes in our center between April 2017 and May 2021 were included in the study. Intervention(s): Women>35 years and PGT cycles with chromosomal structural rearrangements (PGT-SR) were excluded. Embryological and blastocyst ploidy outcomes were compared between SERa+ oocyte, sibling SERa- oocytes and oocytes in SERa- cycles.Main Outcome Measure(s): Embryological outcomes and blastocyst euploidy rate.Results: No significant difference was observed in the normal fertilization rate (82.1 % vs. 77.8 % vs. 83.1 %, respectively, P=0.061), blastocyst formation rate (71.0 % vs. 72.5 % vs. 68.4 %, respectively, P=0.393), good quality blastocyst formation rate (46.4 % vs. 48.3 % vs. 42.6 %, respectively, P=0.198) between the SERa+ oocyte group, sibling SERa- oocyte group and SERa- oocyte group. No significant difference was observed in the euploidy rate (50.0 % vs. 62.5 % vs. 63.3 %, respectively, P=0.324), mosaic rate (12.5 % vs. 9.7 % vs. 13.4 %, respectively, P=0.506) and aneuploidy rate (37.5 % vs. 27.8% vs. 23.2 %, respectively, P=0.137) between the three groups.Conclusion: Our results suggest that the euploidy rate of blastocysts derived from SERa+ cycles and oocytes are not impacted.


2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
G Lópe. Ruiz ◽  
C Olmed. Illueca ◽  
M Bare. Gómez ◽  
S Roy. Bolea ◽  
L Aba. d. Velasco ◽  
...  

Abstract Study question Does Calcymicin improve reproductive outcomes of ICSI cycles in cases of fertilization failure and/or embryo blockage indications? Summary answer The application of the Calcymicin after ICSI improves reproductive outcomes, especially in cases with clinical indication of fertilization failure. What is known already According to the bibliography, deficiencies in the oocyte activation process frequently lead to failed ICSI cycles, and these can be corrected by increasing initial levels of calcium (Ca2+) in the oocyte using assisted oocyte activation techniques (AOA), such as the use of Ca2+ ionophores. Ca2+ Ionophores have been shown to trigger an initial Ca2+ spike in the ooplasm that activates Ca2+/Calmodulin dependent protein kinase II, which initiates the cascade of cellular events leading to oocyte activation. Previous results suggest that Ca2+ ionophore treatment can give live offspring after failed ICSI cycles. Study design, size, duration 270 oocytes collected from 17 patients who presented cycles with low fertilization rates and/or embryo blockage or poor quality embryos (according to ASEBIR’s embryo classification criteria) were retrospectively analyzed. Oocytes were divided into two groups, a control group that underwent conventional IVF/ICSI and another group that underwent an ICSI cycle with AOA. Study groups were defined according to clinical indications and subgroups according to AOA or control. All data were collected from 2017 until 2020. Participants/materials, setting, methods Among the 270 oocytes of the study sample, 142 belonged to the control group and 128 belonged to the AOA group. The AOA group oocytes were activated for 15 minutes immediately after ICSI using a prepared solution containing the Ca2+ ionophore A23187, CultActive© (Gynemed, Germany). Fertilization rate and type, blastocyst formation rate, blastocyst quality, embryo kinetics, and pregnancy rates were analyzed, all of them were compared to FIV/ICSI cycles without oocyte activation (control group). Main results and the role of chance In the analyses of the whole sample of oocytes, the AOA treatment gave a fertilization rate of 72.5%, which was significantly higher compared to 53.8% of the control cycles (p = 0.002). Good quality blastocysts and pregnancy rates were also significantly higher than the control (p = 0.01). In the group with an indication of fertilization failure, a significantly higher fertilization rate was recorded compared to the control (65% and 33%, respectively). A higher rate of abnormal embryos with three pronuclei was also found compared to the control (p &lt; 0.001). There were no significant differences in blastocyst formation rates, quality, or embryo kinetics (p &gt; 0.05). In the group with an indication of embryo blockage/poor embryo quality, a significantly higher rate of good quality blastocysts and lower blastulation time were recorded compared to the control (p &lt; 0.05). Limitations, reasons for caution The safety of the AOA technique with Ca2+ ionophore has not been fully demonstrated. In our study, none of the newborns had malformations, and gestational weeks and birth weights were normal. However, further studies on the safety of this technique are needed to implement it routinely in human reproduction clinics. Wider implications of the findings: According to these findings, an increase in the initial levels of calcium in the oocyte through the application of the Ca2+ ionophore A23187 after ICSI improves the results of failed assisted reproduction cycles, especially in the case of those diagnosed with fertility failure, which is a clear indication for AOA. Trial registration number Not applicable


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