Cloned cattle derived from a novel zona-free embryo reconstruction system

As the demand for cloned embryos and offspring increases, the need arises for the development of nuclear transfer procedures that are improved in both efficiency and ease of operation. Here, we describe a novel zona-free cloning method that doubles the throughput in cloned bovine embryo production over current procedures and generates viable offspring with the same efficiency. Elements of the procedure include zona-free enucleation without a holding pipette, automated fusion of 5-10 oocyte-donor cell pairs and microdrop in vitro culture. Using this system, zona-free embryos were reconstructed from five independent primary cell lines and cultured either singularly (single-IVC) or as aggregates of three (triple-IVC). Blastocysts of transferable quality were obtained at similar rates from zona-free single-IVC, triple-IVC, and control zona-intact embryos (33%, 25%, and 29%, respectively). In a direct comparison, there was no significant difference in development to live calves at term between single-IVC, triple-IVC, and zona-intact embryos derived from the same adult fibroblast line (10%, 13%, and 15%, respectively). This zona-free cloning method could be straightforward for users of conventional cloning procedures to adopt and may prove a simple, fast, and efficient alternative for nuclear cloning of other species as well

A novel antifungal property for the Bacillus licheniformis ComX pheromone and its possible role in inter-kingdom cross-talk

Quorum sensing molecules (QSMs) regulate, through a chemical communication process, multiple complex systems in bacterial and some fungal populations on the basis of cell density. The bacterial QSMs involved in inter-kingdom cross-talk may exhibit antagonistic activity against fungi. This provides an important opportunity for biocontrol of fungal invasion in plants. It has been shown that cultures of Bacillus spp. inhibit fungal growth. Here, we explore the inhibitory potential of the industrial workhorse Bacillus licheniformis NCIMB-8874 and its QSM (ComX pheromone) on the growth of Aspergillus flavus, a cereal, legume, and nut crop pathogen. Our studies show that ComX filtered extracts from cultures of B. licheniformis can cause a significant reduction in the growth of A. flavus NRRL 3357 and ESP 15 at a concentration as low as 13 μg ml−1. This work evidences, for the first time, the inter-kingdom utility of the bacterial quorum sensing ComX pheromone indicating potential antifungal food security against A. flavus