125 A COMPARISON OF SEQUENTIAL MEDIUM WITH NON-SEQUENTIAL MEDIUM: DO SOME PATIENTS' EMBRYOS CULTURE BETTER IN ONE THAN IN THE OTHER?

2009 ◽  
Vol 21 (1) ◽  
pp. 162 ◽  
Author(s):  
K. O. Pomeroy ◽  
S. Foley ◽  
B. Faber ◽  
D. V. Moffitt ◽  
M. D. Johnson

The development of sequential media is often reported as one of the reasons why human IVF pregnancy rates have increased over the last 10 years. The objective was to determine if a single culture medium (global) is as effective in human embryo culture as a sequential medium (G series) and whether embryos of some patients prefer one medium over the other. Patients (n = 144) were placed into one of three treatment arms. Treatment 1, sequential: culture in G Series medium. Treatment 2, nonsequential: culture in global medium. Treatment 3, split: After fertilization, a patient’s embryos are split between the above two treatments. All patients with embryos from April 2007 to January of 2008 (PGD patients were excluded) were allocated to one of the three treatments. Eggs were collected in and fertilized in vitro Life media (GMOPS and GFert, respectively). In treatment 1, patients’ embryos (n = 346) were cultured in sequential media. In treatment 2, patients’ embryos (n = 482) are cultured in nonsequential media. In treatment 3, embryos (n = 420) from each patient are split between sequential and nonsequential media after rinsing in the treatment medium. Patients whose embryos met the criteria (at least 2 embryos, >6 cells, grade excellent to good) were transferred on day 5. All embryos remaining after embryo transfer were frozen on day 5. A maximum of 2 embryos was transferred per patient. Frequencies were compared with a chi square test. Results: The percentage of cases that achieved transfers on Day 5 (an indication of embryo quality) were 44.0%, 50.0% and 68.4% for sequential, nonsequential and split, respectively. Split was significantly different from the sequential (P < 0.05). The percentage of transferrable blastocysts was 27% for sequential, 43% for nonsequential and 39% for split (difference between sequential and the other two was significant at P < 0.0005). Pregnancy rates and implantation rates were 32%/19.0%, 36%/22.5% and 50%/32.2% for sequential, nonsequential and split. (Only implantation rates between split and sequential were significant P < 0.05). For embryos in the split treatment, development to transferable blastocysts was 37.0% in sequential media v. 41.6% in nonsequential medium (not significant). In conclusion, a single medium (nonsequential) for culture to blastocyst yields results similar to embryos cultured in the more complex sequential medium. There is some evidence that a benefit can be gained by splitting embryos between two culture media to increase implantation rates. Further studies need to be done to see if this will result in increased pregnancy rates.

2010 ◽  
Vol 22 (1) ◽  
pp. 301
Author(s):  
B. G. Moura ◽  
J. Almeida ◽  
F. L. Lima ◽  
G. Balbi ◽  
R. Calmerani ◽  
...  

The aim of the work was to study the effects of year period, technical team, breed, beef cattle and dairy cattle on the pregnancy rates in fresh embryos used in bovine transfer of IVF programs. The study was carried out at the fertilization laboratory In Vitro Nyltta Britto de Carvalho, in partnership with In Vitro Brazil, located at the Boa Vista farm, Barra do Pirai, during August 2007 to September 2008, seeking subsidies to improve the use of the technique in the field. During that period, aspirations and inovulations in 3 different periods I (August to December), II (January to April), and III (May to September) were carried out. The jobs were accomplished by 9 technical teams (A, B, C, D, E, F, G, H, and I) rendering services to the laboratory, by working with 2 beef breeds (Brahman and Nelore) and 3 dairy breeds (Gir, Girolando, and Holstein). The different breed receivers were synchronized, and in general, from 6 to 8 days after heat, they received embryo transfer, the cervical way, under low epidural anesthesia, where each female received 1 fresh embryo of IVF. All cows were submitted to gestation diagnosis by rectal palpation and ultrasonography, in general, 42 days after embryo transfer. The numbers of embryo transferred and pregnancy rates were submitted to the chi-square test, which presented significant differences (P < 0.05). There were pregnancy rates of 36.25%a (n = 960), 39.83%a (n = 1180), and 32.59%b (n = 919) in the I, II, and III periods, respectively. Among the 9 technical teams, there were verified pregnancy rates (%) of 33.51d (n = 1313), 30.30d (n = 330), 35.00cd (n = 405), 39.24cd (n = 1060), 59.25a (n = 7), 33.33d (n = 24), 53.57bc (n = 28), 43.31c (n = 157), and 58.33ab (n = 12) for A, B, C, D, E, F, G, H, and I teams, respectively. Among breeds there were rates (%) of 36.89ab (n = 412), 34.68b (n = 1286), 35.13ab (n = 74), 38.94a (n = 1140), and 37.80ab (n = 82) for Brahman, Nelore, Gir, Girolando, and Holstein, respectively. In the study, pregnancy rates (%) of 35.21b (n = 1698) in beef cattle and 38.65a (n = 1296) in dairy cattle were observed. The differences in pregnancy rates with respect to the evaluated factors, may be explained by individual, breed, and nutritional variations of the animals. There are few data in the literature with results on the embryo transfer use of IVF bovine under field conditions.


2006 ◽  
Vol 18 (2) ◽  
pp. 202 ◽  
Author(s):  
O. Dochi ◽  
M. Tanisawa ◽  
S. Goda ◽  
H. Koyama

Repeat-breeding is one of the important factors that affect dairy management. The objective of this study was to investigate the effect of transfer of frozen–thawed IVF embryos on pregnancy in repeat-breeder Holstein cattle. Cumulus–oocyte complexes (COCs) were collected by aspiration of 2–1-mm follicles from ovaries obtained at a local abattoir. COCs were matured for 20 h in TCM-199 supplemented with 5% calf serum (CS) and 0.02 mg/mL of FSH at 38.5°C under a 5% CO2 atmosphere in air. Matured oocytes were inseminated with spermatozoa of 5 × 106/mL in BO solution (Brackett and Oliphant 1975 Biol. Reprod. 12, 260–274) containing 10 mM hypotaurine and 4 units/mL heparin. After 18 h of gamete co-culture, presumptive zygotes were cultured in CR1aa (Rosenkrans et al. 1991 Theriogenology 35, 266) supplemented with 5% CS for 8 days at 38.5°C under 5% CO2, 5% O2, 90% N2 atmosphere in air. After in vitro fertilization, Day 7 and Day 8 blastocysts were frozen in 1.5 M ethylene glycol (EG) in Dulbecco's PBS (DPBS) supplemented with 0.1 M sucrose and 20% CS. Embryos were transferred into a freezing medium, loaded into 0.25-mL straws, and allowed to stand for 15–20 min for equilibration. The straws were then plunged into a −7°C methanol bath of a programmable freezer for 1 min, seeded at −7°C, maintained at −7°C for 15 min, cooled to −30°C at the rate of −0.3°C/min, and then plunged into liquid nitrogen. Recipient animals (43 heifers, 131 cows) included those that did not conceive after being artificially inseminated (AI) 3 to 15 times. The frozen–thawed IVF embryos were directly transferred to the recipient animals 7 days after estrus or AI. Pregnancy rates were analyzed by chi-square test. The results are presented in Table 1. There were no significant differences in the pregnancy rates between treatments. However, a slightly higher pregnancy rate was achieved by embryo transfer after AI. These results suggest that embryo transfer may increase the pregnancy rate in repeat-breeder Holstein cattle. Table 1. Pregnancy rates after transfer of IVF frozen–thawed embryos in repeat-breeder Holstein cattle


2007 ◽  
Vol 19 (1) ◽  
pp. 297
Author(s):  
S. Li ◽  
W. Yu ◽  
J. Fu ◽  
Y. Bai ◽  
F. Jin ◽  
...  

Data collected from commercial embryo transfer programs in 63 farms in China during June 2002 to December 2005 was analyzed to examine the effects of various factors (biopsy, freezing, sample size, embryo development and quality, in vitro culture, and recipient quality) on pregnancy rates of in vivo-biopsied embryos. Embryos were flushed from superovulated dairy cattle and subjected to a biopsy for sexing determination using protocols and sexing kits supplied by AB Technology Ltd. Fresh embryos were implanted on the same day or frozen with AG freeze medium (AB Technology Ltd., Pullman, WA, USA) for later transfer. Recipients were synchronized with CIDA + PG protocols. Embryos were cultured in 6-well dishes containing 1.3 mL of holding medium (AB Technology Ltd.) in each well at room temperature (20–25�C) for examination of embryo survival in vitro. The chi-square test was used in statistic analysis. The implantation of fresh embryos after biopsy did not affect pregnancy rates (49.6%, 257/518) compared to that of non-biopsied fresh and frozen–thawed embryo groups (52.9%, 47/140 and 46.6%, 177/380, respectively). However, for biopsied embryos subjected to frozen and thawed procedures before implantation, particularly for those subjected to the removal of a larger biopsy, a reduced pregnancy rate was observed (41.8%, 297/710; P &lt; 0.01). Pregnancy rates among biopsied embryos at 3 different development stages (morula-early blastocyst, blastocyst, and expanded blastocyst) were not different. Similar results were found between embryo groups of grade 1 and 2. A significant decrease in pregnancy rate (0/10) was observed with embryos held in vitro for a longer period of time (&gt;5 h), suggesting detrimental effects of in vitro conditions on embryo survival. The highest pregnancy rate (68.0%) was observed in recipients synchronized for the first time before being implanted with biopsied embryos. Significant decreases in such rates were found in recipients synchronized for the second or third times or those with an abortion history at the first or second synchronization-implantation treatment (P &lt; 0.01). Better pregnancy rates (45.6%, 41/90; 46.1%, 76/165; and 45.5%, 5/11) were obtained for recipients implanted with biopsied embryos at Days 7.5, 8.0, and 8.5 post-heat detection, respectively, compared to 16% at Day 7 (3/18, P &lt; 0.05). It is concluded that mechanical treatment (cutting) does not reduce the survival of biopsied embryos; however, cryopreservation reduces their ability to survive in vivo. The analyses also suggest that holding embryos in vitro should not be longer than 5 h unless more favorable in vitro conditions can be provided. To achieve better results of implantation of biopsied embryos, embryo transfer should be performed during 7.5–8.5 days post-estrus, and the healthy recipients synchronized for the first time should be used.


2017 ◽  
Vol 29 (1) ◽  
pp. 177
Author(s):  
S. Sato ◽  
O. Dochi ◽  
K. Imai

Reactive oxygen species (ROS) are the main causes of cell damage in bovine embryos in vitro. Folic acid (FA) is an antioxidant that protects cells from ROS. We studied the effect of the addition of FA to maturation and culture media on development of bovine blastocysts and their survival rate after freeze-thawing. Cell-oocyte complexes (COC) were allowed to mature in HEPES (25 mM)-buffered TCM199 (TCM199) supplemented with 5% calf serum (CS), 0.02 AU mL−1 of FSH, and FA (0, 2.5, 25, and 50 µM) for 20 hours (20–25 COC/100-µL droplet of the medium). After 6 hours of gamete co-culture (5 × 106 sperm/mL), presumptive zygotes were cultured in CR1aa medium supplemented with 5% CS and FA (0, 2.5, 25, and 50 μM) for 9 days (day of fertilization = Day 0). Expanded blastocysts that developed from Day 7 to 9 were frozen for further study. Each embryo was frozen in Dulbecco’s PBS (D-PBS) supplemented with 20% CS, 1.5 M ethylene glycol (EG), and 0.1 M sucrose (SUC). Embryos were equilibrated with their freezing medium for 15 min and loaded individually into a 0.25-mL straw. These straws were put into the cooling chamber of a programmable freezer precooled at −7°C. After 2 min, straws were seeded and held for 13 min at −7°C. Next, straws were cooled to −30°C at −0.3°C/min before being plunged into liquid nitrogen. Frozen embryos were thawed by allowing straws to stand in air for 7 s and warming them in a 30°C water bath for 20 s. Thawed embryos were washed twice with D-PBS supplemented with 20% fetal calf serum (FCS), which was warmed to 38°C. They were immersed into the same medium at 38°C for 10 min, and each embryo was cultured in a 20-μL droplet of TCM199 supplemented with 10% FCS and 0.1 mM β-mercaptoethanol (TCM-199-βME) for 72 h. Embryo cleavage rate was observed at 55 h post-insemination. Blastocyst rates were analysed at 9 days post-insemination. Rates of embryos developing into reexpanded, hatching, and hatched blastocyst stages were determined after 72 h of thawing. All data were analysed by the chi-square test and Fisher’s exact test. Cleavage and blastocyst rates after insemination at 55 hours and 9 days, respectively, were not significantly different among media containing 0 μM (n = 278; 74.1% and 39.9%), 2.5 μM (n = 260; 74.2% and 45.8%), 25 μM (n = 258; 75.6% and 45.7%), and 50 μM (n = 253; 76.3% and 42.7%) FA. Survival and hatching rates of frozen and thawed expanded blastocysts after 72 h in culture were 62.5% and 56.3%, respectively, in 0 μM FA (n = 16); 85.2% and 74.1% in 2.5 μM FA (n = 27); 66.7% and 62.5% in 25 μM FA (n = 24); and 68.0% and 64.0% in 50 μM FA (n = 25). Blastocysts cultured in media containing 2.5 μM FA tended to have a higher survival rate than those cultured in media containing 0 μM FA, although this difference was not significant (P = 0.09). Inclusion of FA did not appear to influence development or post-thaw survival of bovine blastocysts produced in vitro.


2010 ◽  
Vol 22 (1) ◽  
pp. 298
Author(s):  
A. C. B. Mariani ◽  
J. F. Souza ◽  
F. B. Lopes ◽  
J. A. T. Souza ◽  
J. L. Ferreira ◽  
...  

The in vitro production (IVP) of bovine embryos is a biotechnique that has created new perspectives in animal reproduction and improvement. However, one of the main issues in the diffusion of this technology concerns the rate of pregnancy of the recipient cows. The objective was to verify the relationship between the fertilization in vitro, the day the recipient female was in estrus, and the pregnancy rates. Two hundred sixty crossbred recipient cows were selected from July to November 2008, synchronized with 2 different protocols at a random moment of the estrous cycle. One of the groups (G1) composed of 110 females that received 2.0 mL of D-cloprostenol, i.m. in only dose. The other group (G2), with 150 females, received a norgestomet auricular implant and 2.0 mg of estradiol benzoate i.m. On the fifth day, the females received, i.m. 200 IU of equine serum gonadotrophin and 150 μg of D-cloprostenol. The implant was removed on the eighth day, and 24 h later all animals received 1.0 mg of estradiol benzoate. The recipients entered in estrus 1 day before, 1 day after, and on the day of IVF. The embryos were transferred on Day 7 after fertilization, trancervically in the ipsilateral uterine horn that contained corpus luteum. The gestation diagnosis was made by ultrasound scan 30 days after the transfer. Data were analyzed using SAS software (version 9.0, SAS Institute, Cary, NC, USA). Initially, data were analyzed by normality test using Proc Univariate; however, a normal distribution was not observed. Data were then analyzed by PROC FREQ with nonparametric method of chi-square test and a significance level of 5%. Among the recipients, 40 were observed in estrus 1 day before IVF (D - 1), 137 on the IVF day (D0) and 83 were observed in estrus 1 day after (D + 1), resulting in a total rate of pregnancy of 38.8%. For each day the resulting pregnancy rates were 42.50% in D - 1, 41.61% in D - 0, and 30.12% in D + 1. These results suggest that there is no relationship between the day of the recipient’s estrus, the day of IVF, and pregnancy rates. There was no influence of the day of estrus in relation to the day of IVF in the gestation rates of the recipients of IVP embryos. However, more studies concerning the recipients are necessary, because they assume an important role in the success of the IVP. The authors thank Brio Genetic and Technology.


2020 ◽  
Vol 5 (3) ◽  
pp. 875
Author(s):  
Asfri Sri Rahmadeni

<p class="Normal1"><em>Commitment is the attitude or behavior of likes or dislikes shown by someone against the organization at work. Indonesia is recorded as a nurse who has a low organizational commitment (76%) so it harms services. This phenomenon in the hospital can be seen from the indicator of the high turnover rate of nurses which reaches 30.9% which exceeds the standard &lt;5%. Many factors affect organizational commitment. The purpose of this study was to analyze the Personal Factors Associated with Organizational Commitment of Hospital Nurses. This study is a cross-sectional analytic study and purposive sampling technique with a sample of 59 nurses. Chi-Square test results obtained personal factors that have a relationship with organizational commitment Nurse Hospital is age with p = 0,000 and education with p = 0,000, personal factors not related to organizational commitment are gender p = 1,053 and years of service with p = 1,147. Hospitals should conduct research/surveys on Organizational Commitment periodically as an evaluation material in the context of the organization's development and development in the future</em>.</p>


Author(s):  
Y. Widyastuti Y. Widyastuti

  ABSTRACT [In advanced countries the number of permanent contraception option ( kontap ) approximately 10 % of all national family planning methods , while in Indonesia reached approximately 5.7 % . Based on data obtained from medical records Bhayangkara Hospital Palembang in 2010 the number of achievement when seen from the percentage of contraceptive use is only 9.10 % The purpose of this study is known age and parity relationship with the selection of contraceptive methods Surgery Women ( MOW ) in Palembang RS.Bhayangkara obstetrics clinic in 2012 . This study uses the analytic survey with cross sectional approach . The population in this study were women who become new acceptors were recorded in medical records at the hospital . Palembang Bhayangkara Hospitals January 1, 2012 to December 31 in 2012 . The sample in this study is part of a whole number of new acceptors ( either using contraception methods MOW and other family planning methods ) were registered in the hospital . Palembang Police Hospitals of January 1, 2012 to December 31 years of data 2012.Analisa univariate and bivariate statistical Chi - Square test with significance level α = 0.05 . The results showed there were 80 respondents ( 46.3 % ) who became MOW contraceptive acceptors , acceptors who have old age by 45 respondents ( 56.3 % ) and who has a young age by 35 respondents ( 43.8 % ) . acceptors which have high parity were 44 respondents ( 55 % ) who had low parity and a total of 36 respondents ( 45 % ) . From the results of Chi-square test value obtained ρ = 0.00 is smaller than α = 0.05 . There is a significant correlation between age and the selection of MOW Chi square test results obtained ρ value = 0.00 and there was a significant association between parity with the election MOW Chi square test results obtained ρ value = 0 , . From these results , it is expected to power health, especially family planning services at the hospital . Palembang Police Hospitals in order to improve the quality of family planning services in an effective, efficient , and safe are considered particularly suitable for users of contraception OperatingMethodWomen(MOW).                                      ABSTRAK Di negara sudah maju jumlah pilihan kontrasepsi mantap (kontap) sekitar 10% dari semua metode KB nasional, sedangkan di Indonesia baru mencapai sekitar 5,7 %. Berdasarkan data yang diperoleh dari Rekam medik  Rumah Sakit Bhayangkara Palembang tahun 2010 Jumlah pencapain tersebut jika dilihat dari persentasi penggunaan kontrasepsi MOW hanya  9,10% Tujuan penelitian ini adalah diketahuinya hubungan umur dan paritas dengan pemilihan alat kontrasepsi Metode Operasi Wanita (MOW) di poliklinik kebidanan RS.Bhayangkara Palembang Tahun 2012. Penelitian ini menggunakan metode survey analitik dengan pendekatan cross sectional. Populasi dalam penelitian ini adalah wanita yang menjadi akseptor KB Baru yang tercatat di rekam medik  di RS. Bhayangkara Palembang tanggal 1 januari 2012 sampai dengan 31 Desember  tahun 2012. Sampel dalam penelitian ini adalah sebagian dari seluruh jumlah akseptor KB Baru (baik yang menggunakan alat kontrasepsi Metode MOW maupun Metode KB lain) yang teregister di RS. Bhayangkara Palembang dari tanggal 1 januari 2012 sampai dengan 31 Desember  tahun 2012.Analisa data dilakukan secara univariat dan bivariat dengan uji statistik Chi-Square dengan tingkat kemaknaan α = 0,05. Hasil penelitian menunjukkan dari 80 responden terdapat (46,3%) yang menjadi akseptor kontrasepsi MOW, akseptor yang memiliki umur tua sebanyak 45 responden (56,3%) dan yang memiliki  umur muda sebanyak 35 responden (43,8%). akseptor yang memiliki paritas tinggi sebanyak 44 responden  (55%) dan yang memiliki  paritas rendah sebanyak 36 responden (45%). Dari hasil uji Chi square didapatkan ρ value = 0,00 lebih kecil dari α = 0,05. Ada hubungan yang bermakna antara umur dengan pemilihan MOW dari hasil uji Chi square didapatkan ρ value = 0,00 dan ada hubungan yang bermakna antara paritas dengan pemilihan MOW hasil uji Chi square didapatkan ρ value = 0,.Dari hasil penelitian ini, diharapkan kepada tenaga kesehatan khususnya pelayanan KB  di RS. Bhayangkara Palembang  agar dapat meningkatkan kualitas pelayanan KB secara efektif, efisien, dan aman yang dianggap cocok bagi penggunanya khususnya mengenai kontrasepsi Metode Operasi Wanita (MOW).    


2020 ◽  
Vol 98 (Supplement_2) ◽  
pp. 2-3
Author(s):  
Theisy P Acosta Pérez

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.


2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Mehdi Dastorani ◽  
Behnam Malekpour ◽  
Mohsen AminSobhani ◽  
Mohammadsadegh Alemrajabi ◽  
Arezoo Mahdian ◽  
...  

Abstract Background Bacterial microleakage is an important cause of apical periodontitis and endodontic treatment failure. This study aimed to assess the bacterial microleakage of nano-mineral trioxide aggregate (nano-MTA) as a sealer, Endoseal MTA, and GuttaFlow Bioseal sealers in atmospheric pressure, and simulated underwater diving and aviation conditions. Methods In this in vitro, experimental study, 180 extracted single-rooted teeth were cleaned and shaped, and were then randomly divided into three groups for single-cone obturation using Endoseal MTA, GuttaFlow Bioseal, or nano-MTA as a sealer. Each group was then randomly divided into three subgroups, and subjected to ambient atmospheric pressure, 2 atm pressure (to simulate underwater diving), and 0.5 atm pressure (to simulate aviation) using a custom-made pressure chamber. The teeth then underwent microbial leakage test using Streptococcus mutans (S. mutans), and the percentage of samples showing microleakage was recorded for up to 1 month, and analyzed using the Chi-square test. Results The three sealer groups were significantly different regarding bacterial microleakage (P < 0.05). The nano-MTA group showed significantly higher microleakage after 15 days than the other two groups (P = 0.006). The effect of pressure on bacterial microleakage was not significant in any sealer group (P > 0.05). Conclusion Within the limitations of this in vitro study, it may be concluded that single-cone obturation technique using nano-MTA as a sealer results in lower resistance to bacterial microleakage compared with the use of GuttaFlow Bioseal, and Endoseal MTA. Pressure changes in simulated underwater diving and aviation conditions had no significant effect on bacterial microleakage. Trial Registration Number This is not a human subject research.


2005 ◽  
Vol 17 (2) ◽  
pp. 276 ◽  
Author(s):  
J. Pryor ◽  
S. Romo ◽  
D.D. Varner ◽  
K. Hinrichs ◽  
C.R. Looney

In commercial bovine in vitro fertilization (IVF) companies, there is a continuous need to improve results. Efforts to maximize in vitro embryo production have included modifications in the use of sperm separation gradients. The development of commercially available sperm centrifugation gradients represents a new possibility of increasing the number of viable sperm that can be obtained from low concentration (fresh or frozen, sexed or unsexed) semen samples in order to improve the efficiency of the IVF system to make embryo production as efficient as possible. The objective of this study was to compare two different separation gradients, as follows: Group 1: Percoll (Sigma, St. Louis, MO, USA), in 45% and 90% gradients; Group 2: EquiPure (Nidacon, Gathenburg, Sweden), in top and bottom layers. Before and after separation, sperm were evaluated at 200× magnification for total motility, and then stained to assess viability at 400× with fast-green/eosin stain (Sigma). Sperm separation was performed using frozen/thawed semen from one bull. Semen was separated by centrifugation at 200g for 30 min in both density gradients. Results obtained from Groups 1 and 2 were compared by chi-square test. Sperm separation with Percoll yielded lower numbers of sperm (average sperm concentration after separation of 92 × 106, vs. 159 × 106 sperm/mL for EquiPure; P < 0.05) but resulted in higher motility (60% vs. 39%, respectively; P < 0.05) of separated sperm. Rates of live sperm cells were not significantly different between groups (69.5% vs. 70%, respectively; P > 0.1). These results indicate that the commercial separation medium EquiPure may be associated with higher sperm concentration levels but with lowered sperm motility when compared to Percoll for bovine sperm separation. However, Equipure provided similar percentages of live sperm when compared to Percoll, which is currently used in our laboratory.


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