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

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.

2005 ◽  
Vol 17 (2) ◽  
pp. 273 ◽  
Author(s):  
Y. Huang ◽  
X. Zhang ◽  
B. Gasparrini ◽  
G.A. Presicce

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.


2004 ◽  
Vol 16 (4) ◽  
pp. 437 ◽  
Author(s):  
Yves Cognié ◽  
Nati Poulin ◽  
Yann Locatelli ◽  
Pascal Mermillod

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.


2013 ◽  
Vol 79 (5) ◽  
pp. 778-783 ◽  
Author(s):  
L.U. Ohlweiler ◽  
D.S. Brum ◽  
F.G. Leivas ◽  
A.B. Moyses ◽  
R.S. Ramos ◽  
...  

Animals ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 3319
Author(s):  
Morgan F. Orsolini ◽  
Stuart A. Meyers ◽  
Pouya Dini

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.


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 ◽  
...  

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.


Animals ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 216
Author(s):  
Hernan Baldassarre

The potential of laparoscopic ovum pick-up (LOPU) followed by in vitro embryo production (IVEP) as a tool for accelerated genetic programs in ruminants is reviewed in this article. In sheep and goats, the LOPU-IVEP platform offers the possibility of producing more offspring from elite females, as the procedure is minimally invasive and can be repeated more times and more frequently in the same animals compared with conventional surgical embryo recovery. On average, ~10 and ~14 viable oocytes are recovered by LOPU from sheep and goats, respectively, which results in 3–5 transferable embryos and >50% pregnancy rate after transfer. LOPU-IVEP has also been applied to prepubertal ruminants of 2–6 months of age, including bovine and buffalo calves. In dairy cattle, the technology has gained momentum in the past few years stemming from the development of genetic marker selection that has allowed predicting the production phenotype of dairy females from shortly after birth. In Holstein calves, we obtained an average of ~22 viable oocytes and ~20% transferable blastocyst rate, followed by >50% pregnancy rate after transfer, declaring the platform ready for commercial application. The present and future of this technology are discussed with a focus on improvements and research needed.


2012 ◽  
Vol 32 (7) ◽  
pp. 409 ◽  
Author(s):  
C. Herrera ◽  
P. Dufourq ◽  
M. Freije ◽  
I. Morikawa ◽  
J.E. Centeno ◽  
...  

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