Lipid Enriched Reduced Nutrient Culture Medium Improves Bovine Blastocyst Formation.

The refinement of embryo culture media is essential in improving embryo viability and in vitro production efficiency. Our previous work demonstrated that the nutrients (carbohydrates, amino acids, and vitamins) in traditional culture media far exceed the need for an embryo and producing developmentally competent embryos in a reduced nutrient environment is feasible. Here, we aim to evaluate the impact of exogenous lipid and L-carnitine supplementation on bovine blastocyst development and refine our RN condition further.

Characterisation of sperm production and morphology in the male Philippine crocodile via voluntary behavioural training.

Philippine crocodiles Crocodylus mindorensis are critically endangered due to agricultural and fishing threats that have severely fragmented their habitat and population in the Philippines. Captive management plans are important to safeguard against their extinction, but the current population in US zoos is small, and breeding is hampered by the slow growth of this species and the danger of introducing differently sized animals for breeding. There is little information regarding the sperm characteristics of crocodilians, and none for Philippine crocodiles.

Nutrient intake and influence on markers of oxidative stress in zoo-managed male snow leopards ().

Oxidative stress (OS) results from the overproduction of reactive species. Nutrient intake can contribute positively or negatively to OS, and the lack of established nutrient requirements for most of the exotic species managed in zoos exacerbates the possibilities for nutrient imbalances that potentially could lead to reactive species production. The objective of this study was to evaluate the influence of nutrient intake and nutritional husbandry on markers of OS in male snow leopards ( = 14) maintained in U.

Fatty acids present in commercial albumin preparations differentially affect development of murine embryos before and during implantation.

Objective: To characterize fatty acid (FA) profile of commercially available albumin products and determine their effect on embryonic development.

Design: Research study.

Setting: Private research facility.

Developmental and molecular response of bovine embryos to reduced nutrients .

Unlabelled: Recent studies in our laboratory have indicated that bovine embryos only use a small amount of the nutrients available to them in culture. Our objective was to evaluate the developmental and molecular response of bovine embryos when nutrient concentrations in the culture medium were significantly reduced. Following IVM and IVF, embryos were cultured in media containing 75, 50, and 25% (experiment 1) or 25, 12.

Factors affecting reproductive traits in male snow leopards ().

Abstract: The population of snow leopards () maintained in US zoos is no longer sustainable due to poor reproductive success. Our objective was to assess reproductive traits in male snow leopards and identify factors (markers of oxidative stress in seminal fluid, surveys of husbandry practices, gonadal and adrenocortical activity, dietary intake of various nutrients, and genetics) that may affect ejaculate traits and subsequent fertility. Ejaculates (2.

Cross-species efficacy of a chemically-defined, soy lecithin-based cryomedium for semen banking in imperiled wild felids.

Felid semen has historically been frozen using an egg yolk-based cryopreservation medium. However, the use of egg introduces several potential concerns, such as variability in composition, microbial contamination, and regulatory issues. In the present study, our aim was to compare a chemically-defined, soy-based medium (SOY) to a commercial egg yolk-based medium (TEY) for the cryopreservation of sperm in four imperiled small cat species.

The landscape of accessible chromatin in bovine oocytes and early embryos.

Chromatin reorganization governs the regulation of gene expression during preimplantation development. However, the landscape of chromatin dynamics in this period has not been explored in bovine. In this study, we constructed a genome-wide map of accessible chromatin in bovine oocytes and early embryos using an improved assay for transposase-accessible chromatin with high-throughput sequencing (ATAC-seq) which revealed unique features of the accessible chromatin during bovine early embryo development.

A novel culture medium with reduced nutrient concentrations supports the development and viability of mouse embryos.

Further refinement of culture media is needed to improve the quality of embryos generated in vitro. Previous results from our laboratory demonstrated that uptake of nutrients by the embryo is significantly less than what is supplied in traditional culture media. Our objective was to determine the impact of reduced nutrient concentrations in culture medium on mouse embryo development, metabolism, and quality as a possible platform for next generation medium formulation.

In Vitro Culture of Embryos from Domestic Cats.

A variety of protocols are available for the production of feline blastocysts from oocytes that are matured and fertilized in vitro. However, most of these protocols utilize media that were developed for other cell types or the embryos of other species. The protocol outlined in this chapter is based on a series of studies that led to the development of feline optimized culture medium (FOCM), a feline-specific, sequential medium system.

Assisted reproductive technologies for endangered species conservation: developing sophisticated protocols with limited access to animals with unique reproductive mechanisms.

Assisted reproductive technologies (ARTs) have been proposed as a means of overcoming the significant challenges of managing small, isolated populations of endangered species in zoos. However, efficient protocols for ARTs do not exist for most endangered species. This review will focus on research efforts to characterize unique reproductive mechanisms and develop species-specific ARTs.

A carnivore embryo's perspective on essential amino acids and ammonium in culture medium: effects on the development of feline embryos.

Carnivores are an interesting model for studies of embryonic amino acid metabolism and ammonium (NH4+) toxicity given the high-protein content of their diets. Our objectives were to examine concentration- and stage-specific effects of essential amino acids (EAA; 0×, 0.125×, 0.

Exogenous growth factors do not affect the development of individually cultured murine embryos.

Purpose: The objective of this study was to evaluate the effects of multiple growth factors on the development of individually cultured murine embryos.

Methods: Embryos produced by in vitro fertilization using in vitro (IVM) or in vivo (IVO) matured oocytes from three strains of mice (CF1, Swiss Webster, B6D2F1) were cultured individually (10 μl) in the absence (control) or presence of growth factors (paf, epidermal growth factor [EGF], insulin-like growth factor 1 [IGF-1], and granulocyte-macrophage colony-stimulating factor [GM-CSF]). Blastocyst formation, hatching, and blastocyst cell numbers (trophectoderm, inner cell mass, and total) were evaluated on days 4 and 5 of culture.

Transporting cumulus complexes using novel meiotic arresting conditions permits maintenance of oocyte developmental competence.

Purpose: The aim of this study is to evaluate the effect of a novel bovine cumulus oocyte complex (COC) shipping media designed to arrest meiotic resumption during transport on meiotic arrest, as well as meiotic resumption, subsequent embryonic development, and embryo quality.

Methods: Bovine cumulus oocyte complexes were transported overnight from the collection facility to the laboratory. COCs were placed in control in vitro maturation (IVM) or in shipping arrest medium (SAM) containing multiple meiotic inhibitors, and then shipped to our laboratory.

Direct and Osmolarity-Dependent Effects of Glycine on Preimplantation Bovine Embryos.

Concentrations of glycine (Gly) in embryo culture media are often lower (~0.1 mM) than those in oviductal or uterine fluids (≥1.2 mM).

Building a better mouse embryo assay: effects of mouse strain and in vitro maturation on sensitivity to contaminants of the culture environment.

Purpose: The aim of this study is to compare the sensitivity of the standard one-cell mouse embryo assay (MEA) to that using in vitro-matured oocytes from hybrid and outbred mice.

Methods: The study was done by culturing embryos in the presence or absence of two concentrations (0.0005 or 0.

Applying metabolomic analyses to the practice of embryology: physiology, development and assisted reproductive technology.

The advent of metabolomics technology and its application to small samples has allowed us to non-invasively monitor the metabolic activity of embryos in a complex culture environment. The aim of this study was to apply metabolomics technology to the analysis of individual embryos from several species during in vitro development to gain an insight into the metabolomics pathways used by embryos and their relationship with embryo quality. Alanine is produced by both in vivo- and in vitro-derived human, murine, bovine and porcine embryos.

The effects of permeating cryoprotectants on intracellular free-calcium concentrations and developmental potential of in vitro-matured feline oocytes.

Embryos produced from vitrified feline oocytes have resulted in pregnancies, but the efficiency of oocyte vitrification in cats is still low. Our objectives were to evaluate the effects of exposing feline oocytes to ethylene glycol (EG), propanediol (PrOH) and dimethyl sulfoxide (DMSO) on changes in intracellular free-calcium concentrations ([Ca(2+)]i), the time needed for enzymatic digestion of the zona pellucida (ZP), the incidence of parthenogenetic activation and degeneration and embryonic development following in vitro fertilisation (IVF). All of the chemicals tested altered [Ca(2+)]i, but changes in [Ca(2+)]i, resistance of the ZP to enzymatic digestion and the incidence of parthenogenetic activation (<5% for all treatments) were not affected (P>0.

Composition of commercial media used for human embryo culture.

Objective: To determine the composition of commercially available culture media and test whether differences in composition are biologically relevant in a murine model.

Design: Experimental laboratory study.

Setting: University-based laboratory.

Fecal endocrine monitoring of reproduction in female snow leopards (Uncia uncia).

Although the snow leopard (Uncia uncia) is a common endangered felid species in zoos, little is known about the complex endocrine interactions controlling ovarian function and conception in this species. The goal of this work was to characterize ovarian activity throughout the estrous cycle, nonpregnant luteal phase (pseudopregnancy), and gestation in female snow leopards. This goal was accomplished using an enzyme immunoassay to measure fecal concentrations of estrogen metabolites (E) and progesterone metabolites (P).

Reversible meiotic arrest in feline oocytes.

Increasing intracellular concentrations of cyclic adenosine monophosphate (cAMP) within the cumulus-oocyte complex (COC) inhibits or delays spontaneous oocyte maturation and improves the developmental competence of the oocyte in many species, but information for carnivores is limited. The objectives of the present study were to describe the effects of isobutyl methylxanthine (IBMX), which decreases cAMP degradation, and forskolin, which increases cAMP production, on spontaneous and induced maturation (by equine chorionic gonadotrophin (eCG) and epidermal growth factor (EGF)) of feline oocytes and to evaluate the reversibility of IBMX-induced arrest by measuring the resumption of meiosis and embryonic development following IVF. IBMX decreased (P<0.

Analysis of cat oocyte activation methods for the generation of feline disease models by nuclear transfer.

Background: Somatic cell nuclear transfer in cats offers a useful tool for the generation of valuable research models. However, low birth rates after nuclear transfer hamper exploitation of the full potential of the technology. Poor embryo development after activation of the reconstructed oocytes seems to be responsible, at least in part, for the low efficiency.