scholarly journals 245 OOCYTE RECOVERY BY OVUM PICK UP AND EMBRYO PRODUCTION IN MURRAH AND NILI-RAVI BUFFALOES (BUBALUS BUBALIS) IMPORTED IN CHINA

2005 ◽  
Vol 17 (2) ◽  
pp. 273 ◽  
Author(s):  
Y. Huang ◽  
X. Zhang ◽  
B. Gasparrini ◽  
G.A. Presicce
Keyword(s):  
Essential Amino Acids ◽  
Reproductive Technologies ◽  
Meat Production ◽  
Embryo Production ◽  
Vitro Fertilization ◽  
Preliminary Results ◽  
Swamp Buffaloes ◽  
In Vitro Embryo Production ◽  
Humidified Air

In this preliminary study, in vitro embryo production and cryopreservation in two river type buffaloes (Murrah and Nili-Ravi) imported into China have been carried out. The objective of the study was enhancement of the genetic merit and productive performances of imported river buffaloes in conjunction with the utilization of local swamp buffaloes. In order to improve milk and meat production in China local swamp buffaloes (2n = 48), which are the predominant subspecies, have been crossbred with imported river buffaloes (Murrah and Nili-Ravi: 2n = 50). At present, several hundred thousand crossbred heads have been produced, and although both males and females can reproduce with 2n = 49 crossbred buffaloes, their reproductive performances are significantly reduced when compared to 2n = 50 buffaloes. As an alternative approach, a program of embryo production in river buffaloes and transfer into both river and swamp buffaloes has been implemented at the Guangxi Buffalo Research Institute, in Nanning, P.R. China. Some preliminary results are presented: from a start-up experiment, a total of 46 river buffaloes were subjected to 2 to 3 ovum pickup sessions at 4-day intervals. A total of 750 antral follicles were punctured and 495 (66%) cumulus-oocyte complexes (COCs) were retrieved. Only COCs characterized by at least one layer of granulosa cells (n = 451; 91.1%) were considered for in vitro maturation (IVM). COCs were matured in TCM 199 + 10% FCS, 0.5 μg/mL FSH, 5 μg/mL LH, and 1 μg/mL estradiol in the presence of cysteamine (50 μM) at 39°C under 5% CO2 in humidified air for 24 h. Of the initial 451 COCs matured, only 277 could be considered for in vitro fertilization (IVF). IVF was performed at 39°C under CO2 in humidified air in TALP medium supplemented with 0.2 mM penicillamine, 0.1 mM hypotaurine, and 0.01 mM heparin. Frozen/thawed sperm from a tested bull was treated by swim-up procedure and used at a final concentration of 20 million/mL. Following 20 to 22 h of co-incubation, presumptive zygotes were cultured in SOF medium, supplemented with essential and non-essential amino acids and 8 mg/mL BSA, in a gas atmosphere of 5% CO2, 7% O2, and 88% N2. A total of 41 (14.8%) blastocysts were produced, of which 33 were vitrified and 8 transferred immediately into available swamp and river buffalo recipients. Two calves were born (25%) from the transfer of fresh embryos into one river and one swamp buffalo. In vitro embryo production in the buffalo species is still characterized by a high degree of variable results. However, these preliminary results reinforce the need to implement newly developed reproductive technologies not only for speeding up genetic gain of already productive species, but also for the utilization of local breeds characterized by reduced productive performance.

186 SUPPLEMENTATION WITH LOW DOSES OF DIMETHYL SULFOXIDE DURING IN VITRO MATURATION RESULTS IN IMPROVED IN VITRO EMBRYO PRODUCTION IN CATTLE

2017 ◽  
Vol 29 (1) ◽  
pp. 201
Author(s):  
A. E. Ynsaurralde ◽  
M. Suvá ◽  
R. Bevacqua ◽  
S. Munilla ◽  
C. Luchetti ◽  
...  
Keyword(s):  
Dimethyl Sulfoxide ◽  
Embryo Development ◽  
In Vitro Maturation ◽  
Polar Body ◽  
Predictive Equation ◽  
Embryo Production ◽  
Vitro Fertilization ◽  
First Polar Body ◽  
In Vitro Embryo Production

Oocyte in vitro maturation (IVM) is crucial for subsequent in vitro embryo production. It involves acquisition of competence for fertilization and embryo development. Therefore, its optimization could have a direct impact on in vitro embryo development. Dimethyl sulfoxide (DMSO) is commonly used as solvent or vehicle, but also increases the membrane permeability and behaves as a scavenger of cytotoxic free radicals. The aim of this study was to evaluate the effect of DMSO supplementation during bovine oocyte maturation on subsequent in vitro embryo development and to determine the optimal usage dose with no toxic effect. To this aim, cumulus-oocyte complexes were collected from slaughterhouse ovaries and IVM in TCM 199 containing 10% fetal bovine serum, 10 µg mL−1 of FSH, 0.3 mM sodium pyruvate, 100 mM cysteamine, and 2% antibiotic-antimycotic. The oocytes were incubated for 24 h at 6.5% CO2 in humidified air at 38.5°C. For Experiment 1, IVM medium was supplemented with DMSO at concentrations of 0, 0.1, 0.5, 1, or 10% (vol/vol) DMSO (n = 241, 195, 42, 192, 172 oocytes) and IVM rate was determined by presence of the first polar body. For Experiment 2, 0, 0.1, 0.25, 0.5, 0.75, 1, or 10% (vol/vol) DMSO (n = 446, 322, 65, 194, 77, 250, 39 oocytes) was supplemented to IVM medium and cleavage and blastocyst rates were determined to establish the optimal usage dose. In vitro fertilization was performed according to Brackett and Oliphant (1975), with 16 × 106 spermatozoa/mL for 5 h. Afterwards, presumptive zygotes were cultured in SOF for 7 days at 38.5°C and 5% O2. Cleavage and blastocyst rates were determined on Days 2 and 7, respectively. Results were statistically analysed using Fisher’s exact test by GraphPad Prism software (GraphPad Software Inc., La Jolla, CA, USA). Also, the percentage of blastocyst was adjusted to DMSO concentration using the R software quadratic regression model. The optimum usage dose was determined by calculating the maximum of the estimated predictive equation. In vitro maturation in 10% DMSO resulted in significantly lower first polar body extrusion rates (0% = 74%a, 0.1% = 73%a, 0.5% = 83%a, 1% = 66%a, and 10% = 8%b; different letters indicate statistical differences) and lower cleavage rates (0% = 75%a, 0.1% = 77%a, 0.25% = 80%a, 0.5% = 79%a, 0.75% = 78%a, 1% = 77%a, and 10% = 3%b) than the other treatments. Furthermore, blastocyst production was higher for the 0.25 and 0.5% (vol/vol) supplemented DMSO groups (0% = 26%b, 0.1% = 37%ab, 0.25% = 40%a, 0.5% = 41%a, 0.75% = 34%ab, 1% = 23%b, and 10% = 0%c). The predictive equation results indicate that the maximum percentage of blastocysts is obtained with a concentration of 0.458% (vol/vol) of DMSO. In conclusion, DMSO supplementation during IVM of bovine oocytes had a positive effect on in vitro development. Further studies will be carried out to elucidate its mechanism of action.

State-of-the-art production, conservation and transfer of in-vitro-produced embryos in small ruminants

2004 ◽  
Vol 16 (4) ◽  
pp. 437 ◽  
Author(s):  
Yves Cognié ◽  
Nati Poulin ◽  
Yann Locatelli ◽  
Pascal Mermillod
Keyword(s):  
Oocyte Maturation ◽  
Small Ruminants ◽  
Reproductive Technologies ◽  
Defined Medium ◽  
Blastocyst Stage ◽  
Embryo Production ◽  
Oocyte Competence ◽  
Epidermal Growth ◽  
In Vitro Embryo Production

Today, although not efficient enough to replace multiple ovulation and embryo transfer, in vitro embryo production for small ruminants is a platform for new reproductive technologies, such as embryo sexing, transgenesis and cloning. The in vitro embryo-production system developed for sheep and goats is more efficient now than 15 years ago, but could still be improved. Laparoscopic collection of oocytes in live animals treated with gonadotrophin indicates a promising future for the application of this technology to genetic improvement programmes. Oocyte maturation in defined medium with epidermal growth factor and cysteamine appears as efficient as oocyte maturation in follicular fluid-supplemented medium and allows future study of the effect of other factors involved in the cytoplasmic maturation of oocytes from these species. Further efforts have to be made to standardise the semen-capacitating process and to improve the quality and freezability of in-vitro-produced (IVP) embryos. The optimisation of IVP procedures for deer species has required the study of the seasonal variation of oocyte competence and the development of a specific methodology to allow the culture of embryos up to the blastocyst stage.

scholarly journals Evaluation of the sperm separability of blanc-blue-belge bull by swim-up method and in vitro embryo production with hybrid Zebu bovine oocyte in Vietnam

2020 ◽  
Vol 18 (1) ◽  
pp. 59-66
Author(s):  
Nguyen Huu Duc ◽  
Pham Thu Giang ◽  
Tran Thi Binh Nguyen ◽  
Bui Dai Phong
Keyword(s):  
In Vitro Fertilization ◽  
Calf Serum ◽  
Basic Medium ◽  
Embryo Production ◽  
Vitro Fertilization ◽  
Bovine Sperm ◽  
Maturation Medium ◽  
In Vitro Embryo Production ◽  
The Right

The objective of this study was to determine the right conditions for the separation of Blanc-Blue-Belge bovine sperm (BBB) by swim-up mothed; determine the maturity of hybrid Zebu bovine eggs; and culture of embryos after in vitro fertilization. After 60-80 minutes of swim-up in CAP-05, BBB bovine sperms were healthy, straight movement and separated with a concentration of 106 sperm/ml. Hybrid Zebu bovine eggs developed and matured in the maturation medium with the basic medium TCM-199 supplemented with 10% calf serum, FSH (0.75 µg / ml), LH (0.15 µg / ml) and Estradiol (2.5 µg / ml), the results showed that the IVM-08 medium had significantly higher maturation rates than IVM-03, the proportion of mature eggs reached 71,11% compared to 51.69%, respectively (P <0.01). In vitro fertilization of hybrid Zebu bovine egg in IVF-08 medium. In vitro fertilized embryos (BBB x hybrid Zebu) developed from bovine sperms separated by the swim-up method achieved a better rate of morula-blastocysts in IVC-09 than IVC-06 medium, 21.68% compared to 8.56%, respectively (P <0.01). The conclusion was that the suitable conditions for BBB bovine sperm separation and in vitro embryo production (BBB x hybrid Zebu) were determined. This is the premise to create bovine semen, BBB bovine embryos with defined gender.

205 GLUCOSE-6-PHOSPHATE DEHYDROGENAZE ACTIVITY IN OVINE OOCYTES IS ASSOCIATED WITH PRE-IMPLANTATION EMBRYONIC DEVELOPMENT IN VITRO

2011 ◽  
Vol 23 (1) ◽  
pp. 201 ◽  
Author(s):  
F. Asghari ◽  
M. Shahidi ◽  
Y. Chashnidel ◽  
H. Deldar ◽  
Z. Ansari-Pirsaraei ◽  
...  
Keyword(s):  
Oocyte Quality ◽  
G6pd Activity ◽  
Blue Staining ◽  
Control Group ◽  
Embryo Production ◽  
Brilliant Cresyl Blue ◽  
Cresyl Blue ◽  
In Vitro Embryo Production ◽  
Humidified Air

A large proportion of ovine oocytes fail to develop into viable embryos following maturation, fertilization, and culture in vitro. Accurate, fast, and noninvasive predictors of ovine oocyte quality are therefore in urgent need for oocyte selection before in vitro maturation (IVM). Recent studies have shown that oocyte competence can be predicted through the presence of the glucose-6-phosphate dehydrogenase (G6PD) enzyme, as indicated by brilliant cresyl blue (BCB), a dye that can be degraded by G6PD. Thus, oocytes that have completed their growth phase show decreased G6PD activity and exhibit cytoplasm with a blue colouration (BCB+), whereas growing oocytes are expected to have a high level of G6PD, which results in colourless cytoplasm (BCB–). The brilliant cresyl blue staining test, as a noninvasive intrinsic criterion, has been successfully used to identify the more competent oocytes in various species. Therefore, this study aimed to investigate whether BCB staining, as an indicator of G6PD activity, can be used to select developmentally competent ovine oocytes before IVM and thereby increase the efficiency of in vitro embryo production. Ovine ovaries were obtained from a local slaughterhouse and transported to the laboratory, where cumulus–oocyte complexes (COC) were recovered by slicing the ovaries. Only oocytes with one or more complete layers of unexpanded cumulus cells and a homogeneous cytoplasm were used. The COC were exposed to 26 mM BCB diluted in modified Dulbecco’s PBS for 90 min at 39°C in humidified air. After BCB exposure, the COC were examined under a stereomicroscope and divided into 2 groups: BCB+ (blue cytoplasm, low G6PD activity) and BCB– (colourless cytoplasm, high G6PD activity). Cumulus–oocyte complexes in the control group were incubated for IVM directly after selection, without exposure to BCB dye. After IVM, oocytes were subjected to IVF followed by embryo culture for 7 days (5% CO2, 39°C, humidified air). Results were analysed by a chi-square test, and P < 0.05 was considered statistically significant. The proportion of oocytes that cleaved by Day 2 after insemination was significantly (P < 0.05) higher for the control and BCB+ groups [67.3% (68/101) and 71.7% (81/113), respectively] than for the BCB– group [50.5% (46/91)]. Significant differences among groups were also observed on Day 7 after fertilization, when the embryos reached the blastocyst stage of development. The BCB+ group yielded a significantly (P < 0.05) higher proportion of blastocysts [34.5% (39/113)] than both the control [20.8% (21/101)] and BCB– [4.3% (4/93)] groups. In addition, the blastocyst rate of development in the control group was significantly (P < 0.05) higher than that for the BCB– group. In conclusion, results of this study show that selection of ovine oocytes based on G6PD activity through the BCB test can be used as an efficient predictor of in vitro embryonic developmental competence. This positive predictive parameter of oocyte quality may also be useful in increasing the efficiency of blastocyst production during in vitro embryo production procedures in the ovine.

scholarly journals Oocyte aspiration and in vitro embryo production in Jersey cows with selenium-supplemented diet

2012 ◽  
Vol 64 (4) ◽  
pp. 787-795
Author(s):  
G.V. Moraes ◽  
J.R. Azevedo ◽  
T.C. Carneiro ◽  
F.L.B. Cavalieri ◽  
M. Mataveli ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Mineral Salt ◽  
Embryo Production ◽  
Vitro Fertilization ◽  
Standard Procedures ◽  
Daily Diet ◽  
In Vitro Embryo Production ◽  
Oocyte Aspiration ◽  
Jersey Cows

The effects of selenium (Se) in Jersey cows' diet on the aspiration of oocytes and production of embryos in vitro were studied. Groups with five Jersey cows received 3.2mg or 9.6mg Se daily, provided in the feed concentrate. Six follicular aspirations were carried out every 15 days, using only the last 5. The oocytes were classified, and standard procedures were carried out for maturation, fertilization and cultivation. The total number of oocytes (35.11±2.65 vs 23.10±2.16) and degree 1 oocytes (11.61±2.65 vs 4.75±0.97) were higher in the group that received 9.6mg Se and the quantity of naked oocytes (3.23±0.87 vs 6.22±1.18) was lower in this group. The aspirated oocytes from the cows treated with 9.6mg Se/day resulted in higher (P<0.05) embryo production 21.98±2.37 vs 13.12±1.59). No difference was observed in serum Se concentration between the two groups. It is recommended that the daily diet be supplemented with 100g mineral salt containing 9.6mg Se, since this rate rendered a larger production of oocytes, higher quantity of degree 1 oocytes and greater production of embryos in the process of in vitro fertilization.

Update on advanced semen-processing technologies and their application for in vitro embryo production in horses

2019 ◽  
Vol 31 (12) ◽  
pp. 1771
Author(s):  
Lee H. Morris ◽  
Lisa J. Maclellan
Keyword(s):  
Intracytoplasmic Sperm Injection ◽  
Reproductive Technologies ◽  
Breeding Program ◽  
Embryo Production ◽  
Processing Technologies ◽  
Advanced Technologies ◽  
The Status ◽  
In Vitro Embryo Production ◽  
Semen Processing

The increased commercialisation of intracytoplasmic sperm injection (ICSI) in horses creates more opportunities to incorporate advanced reproductive technologies, such as sex-sorted, refrozen and lyophilised spermatozoa, into a breeding program. This paper reviews the status of these semen-handling technologies in light of their use in equine ICSI programs. Pregnancies have been achieved from each of these advanced technologies when combined with ICSI in horses, but refinements in the semen-handling processes underpinning these technologies are currently being explored to produce more reliable and practical improvements in the results from equine ICSI.

scholarly journals An Update on Semen Physiology, Technologies, and Selection Techniques for the Advancement of In Vitro Equine Embryo Production: Section II

Animals ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 3319
Author(s):  
Morgan F. Orsolini ◽  
Stuart A. Meyers ◽  
Pouya Dini
Keyword(s):  
Assisted Reproductive Technologies ◽  
Reproductive Technologies ◽  
Future Application ◽  
Embryo Production ◽  
Future Developments ◽  
Production Section ◽  
In Vitro Embryo Production ◽  
Equine Industry ◽  
Selection Of

As the use of assisted reproductive technologies (ART) and in vitro embryo production (IVP) expand in the equine industry, it has become necessary to further our understanding of available semen selection techniques. This segment of our two-section review will focus on the selection of spermatozoa based on quality and sex for equine intracytoplasmic sperm injection (ICSI), as well as current and future developments in sperm sorting technologies. Ultimately, novel methods of semen selection will be assessed based on their efficacy in other species and their relevance and future application towards ARTs in the horse.

scholarly journals Genetic diversity in oocyte donors used in in vitro bovine embryo production programs in Brazil

2019 ◽  
Vol 32 (2) ◽  
pp. 90-99
Author(s):  
Wilder Hernando Ortiz Vega ◽  
Celia Raquel Quirino ◽  
Aylton Bartholazzi Junior ◽  
Clara Slade Oliveira ◽  
Raquel Varella Serapião ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Microsatellite Markers ◽  
Assisted Reproductive Technologies ◽  
Reproductive Technologies ◽  
Bovine Embryo ◽  
Embryo Production ◽  
Low Genetic Diversity ◽  
In Vitro Embryo Production ◽  
Population Interactions

Background: Current reproductive management of bovine elite populations involves the use of assisted reproductive technologies (ARTs), aiming to obtain the greatest genetic gain. However, inadequate use of ARTs may lead to loss of genetic diversity in the offspring. Objective: To assess the genetic diversity in elite female cattle populations used in commercial in vitro embryo production. Methods: Using genetic and ecological approaches for the study of populations based on microsatellite markers, we assessed the genetic diversity between and within populations of cows used in commercial in vitro embryo production programs in Brazil. Results: Endogamy within populations varied from zero to 9.1%, while heterozygosity between populations (FST) was <0.05 in the different population interactions. AMOVA showed 1% variation between populations, 8% between individuals and 91% within individuals. The dimensionality reduction method utilized indicated a lack of structure in the populations analyzed, identifying two main clusters in the three populations. Conclusions: Low genetic diversity between cow populations associated with commercial programs of in vitro embryo production in Brazil was evidenced. Variable levels of endogamy within the populations were observed. Approaches of population genetics as well as ecological diversity can be implemented to more thoroughly estimate genetic diversity in livestock populations.Keywords: allele frequencies, heterozygosity, inbreeding, microsatellite markers, oocyte. ResumenAntecedentes: El actual manejo reproductivo en poblaciones de bovinos de élite incluye la utilización de tecnologías de reproducción asistida (ARTs) con el fin de obtener mayor ganancia genética. Sin embargo, el uso inadecuado de las ART puede llevar a la pérdida de diversidad genética en los descendientes. Objetivo: Evaluar la diversidad genética en poblaciones de vacas de élite utilizadas en la producción comercial de embriones bovinos in vitro. Métodos: Utilizando abordajes de la genética y ecología de poblaciones basados en marcadores microsatélites, evaluamos la diversidad genética entre y dentro de poblaciones de vacas participantes de programas comerciales de producción de embriones in vitro en Brasil. Resultados: La endogamia dentro de las poblaciones varió de cero a 9.1%, mientras que la heterocigosidad entre poblaciones (FST) fue <0.05 en las diferentes interacciones de la población. El AMOVA mostró variación del 1% entre poblaciones, 8% entre individuos y 91% dentro de individuos. El método de reducción de dimensionalidad utilizado indicó una falta de estructura en las poblaciones analizadas, identificando dos grupos principales en las tres poblaciones. Conclusiones: Se evidenció una baja diversidad genética entre las poblaciones de vacas asociadas a programas comerciales de producción de embriones in vitro en Brasil. Se evidenciaron niveles variables de endogamia entre las poblaciones. Abordajes de la genética poblacional, así como de diversidad ecológica pueden ser implementados para estimar de manera más amplia la diversidad genética en poblaciones animales de interés pecuario.Palabras clave: endogamia, frecuencia alélica, heterozigosidad, marcadores microsatélites, ovocito. ResumoAntecedentes: O atual manejo reprodutivo das populações de elite em bovinos envolve o uso de tecnologias de reprodução assistida (ARTs), visando obter o maior ganho genético. No entanto, o uso inadequado de ARTs pode levar à perda de diversidade genética na prole. Objetivo: Avaliar a diversidade genética em populações de vacas de elite utilizadas na produção comercial de embriões bovinos in vitro. Métodos: Utilizando abordagens da genética e ecologia de populações baseadas em marcadores microssatélites, foi avaliada a diversidade genética entre e dentro das populações de vacas participantes de programas comercias de produção in vitro de embriões. Resultados: A endogamia dentro das populações variou de zero a 9,1%, enquanto a heterozigosidade entre populações (FST) foi <0,05 nas diferentes interações populacionais. AMOVA mostrou variação de 1% entre populações, 8% entre indivíduos e 91% dentro de indivíduos. O método de redução de dimensionalidade utilizado indicou uma falta de estrutura nas populações analisadas, identificando dois clusters principais nas três populações. Conclusões: Baixa diversidade genética entre populações de vacas associadas a programas de produção in vitro de embriões foi evidenciada. Níveis de endogamia variáveis dentro das populações foram observados. Abordagens da genética populacional assim como de diversidade ecológica podem ser implementadas na tentativa de estimar de maneira mais abrangente a diversidade genética em populações animais de interesse pecuário.Palavras–chave: endogamia, frequência alélica, heterozigosidade, marcadores microssatélite, oócito.

scholarly journals 270IN VITRO FERTILIZATION OF BUFFALO (BUBALUS BUBALIS) OOCYTES: EFFECTS OF MEDIA AND SPERM MOTILITY INDUCING AGENTS

2004 ◽  
Vol 16 (2) ◽  
pp. 255 ◽  
Author(s):  
B. Gasparrini ◽  
L. Boccia ◽  
A. De Rosa ◽  
D. Vecchio ◽  
R. Di Palo ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Significant Interaction ◽  
Production Efficiency ◽  
Blastocyst Stage ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Lower Efficiency ◽  
Vitro Fertilization ◽  
Humidified Air

The identification of an optimal in vitro fertilization system is critical in order to improve the in vitro embryo production efficiency in buffalo species. The aim of this work was to evaluate the effects of fertilization media and sperm motility inducing factors (SMIF) on cleavage and blastocyst rates in buffalo species. Cumulus-oocytes complexes (n=516), recovered from slaughtered animals, were matured in vitro in TCM 199+10 % FCS, 0.5μgmL−1 FSH, 5μgmL−1 LH, 1μgmL−1 17β-estradiol and 50μM cysteamine, at 38.5°C under 5% CO2 in humidified air for 24 hours. The mature oocytes were randomly assigned to four groups for fertilization. In particular, IVF was carried out at 38.5°C under 5% CO2 in humidified air in either Tyrode’s modified medium or Brackett Oliphant medium, in the presence of 0.01mM heparin;; each medium was supplemented with either a mixture of 0.2mM penicillamine and 0.1mM hypotaurine or 5mM caffeine. Frozen-thawed sperm from a tested bull was treated by the swim-up procedure and used at a final concentration of 206mL−1. After 20–22h presumptive zygotes were cultured in SOF medium, supplemented with essential and non-essential amino acids and BSA, in a gas atmosphere of 5% CO2, 7% O2 , and 88% N2, up to the blastocyst stage. Cleavage rates and blastocyst yields were analyzed by a full factorial model 2×2 with medium and SMIF effects (SPSS 11.0). The analysis used permits the identification of statistical differences between treatments irrespective of an interaction (Searle SR. 1971. Linear model. Ed. John Wiley &amp; Sons;; XXI:533). The comparison of the two media, irrespective of the SMIF used, did not show any difference in cleavage rate (43.7% v. 39.3%, respectively, in TALP and BO). On the contrary higher cleavage rates were recorded with hypotaurine-penicillamine v. caffeine (47.7% v. 35.3%; P&lt;0.05), regardless of the medium employed. However, a significant interaction between media and SMIF was found;; in fact the addition of hypotaurine and penicillamine significantly improved cleavage rate compared with caffeine in TALP medium (59.6% v. 27.9%; P&lt;0.05) whereas no differences were observed in BO (35.9% v. 42.7%, respectively). With regard to blastocyst yield a significant effect of medium was also found, with the highest embryo production in TALP v. BO (13.9% v. 6.8%; P&lt;0.05). Blastocyst rate was improved in the presence of hypotaurine-penicillamine v. caffeine (13.6% v. 7.2%; P&lt;0.05). Furthermore there was a significant interaction between medium and SMIF, with the highest embryo yields in the presence of hypotaurine-penicillamine v. caffeine in TALP (20.7% v. 7.1%, respectively;; P&lt;0.05) but not in BO (6.4% v. 7.2%, respectively). The differences we found disappeared when the embryo yield was calculated in relation to the cleaved eggs, with the exception of a lower efficiency of BO v. TALP (15.9% v. 31.1%; P=0.057), suggesting an influence of BO also on post-fertilization development.

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