Leveraging a natural murine meiotic drive to suppress invasive populations

Invasive rodents are a major cause of environmental damage and biodiversity loss, particularly on islands. Unlike insects, genetic biocontrol strategies including populationsuppressing gene drives with biased inheritance have not been developed in mice. Here, we demonstrate a gene drive strategy (tCRISPR) that leverages super-Mendelian transmission of the t haplotype to spread inactivating mutations in a haplosufficient female fertility gene (Prl). Using spatially explicit individual-based in silico modeling, we show that tCRISPR can eradicate island populations under a range of realistic field-based parameter values. We also engineer transgenic tCRISPR mice that, crucially, exhibit biased transmission of the modified t haplotype and Prl mutations at levels our modeling predicts would be sufficient for eradication. This is an example of a feasible gene drive system for invasive alien rodent population control.Luke Gierusa, Aysegul Birandc, Mark D. Buntinga, Gelshan I. Godahewa, Sandra G. Piltz Kevin P. Oh, Antoinette J. Piaggio, David W. Threadgill, John Godwin, Owain Edwards, Phillip Cassey, Joshua V. Ross, Thomas A. A. Prowse and Paul Q. Thoma

Feed intake and performance of sheep grazing semiarid grassland in response to different grazing systems

Effects of grazing management systems (GS) on biomass production and nutritional quality of rangeland vegetation in semiarid regions are extensively studied; however, limited information is available regarding their effects on diet digestibility and feed intake of grazing livestock. We therefore analyzed digestibility of ingested organic matter (dOM), organic matter intake (OMI), and live weight gain (LWG) of sheep in a grazing experiment established in the Inner Mongolian steppe of China, where two GS were tested for six different grazing intensities (GI) from very light to heavy grazing. For the continuous grazing system, sheep grazed the same plots each year, and for the alternating system, grazing and hay making were alternated annually between two adjacent plots. In July, August, and September 2009 and 2010, feed intake and live weight of sheep were determined. The GS did not affect dOM (P-=-0.101), OMI (P-=-0.381), and LWG of sheep (P-=-0.701). Across both GS LWG decreased from 98 g-·-d-1 for GI1 to 62 g-·-d-1 for GI6 (P-<-0.001; R2-=-0.42). There were no interactions between GS and GI for all measured parameters (P-≥-0.061), indicating that alternating grazing did not compensate for negative effects of heavy grazing even after 4 yr of grassland use. In summary, our study showed that irrespective of GI, alternating grassland use does not improve dOM, OMI, and hence, LWG of sheep. However, it might enhance revenues and ecological sustainability in the long term when compared to the common practice of continuous grazing at very high stocking rates. © 2014 The Society for Range Management.The Rangeland Ecology & Management archives are made available by the Society for Range Management and the University of Arizona Libraries. Contact [email protected] for further information

Bacteriocin-like inhibitory substance of Pediococcus pentosaceus as a biopreservative for Listeria sp. control in ready-to-eat pork ham

The growing demand of consumers for synthetic chemical-free foods has increased the search for natural preservatives such as bacteriocins and bacteriocin-like inhibitory substances (BLIS) to give them adequate microbiological safety, sensory characteristics, and shelf life. In this study, the antimicrobial activity of BLIS produced by Pediococcus pentosaceus ATCC 43200 was compared with that of nisin. Lactobacillus sakei ATCC 15521, Listeria seeligeri NCTC 11289, Enterococcus En2052 and En2865, and Listeria monocytogenes CECT 934 and NADC 2045 exhibited larger inhibition halos in BLIS-treated than in Nisaplin-treated samples, unlike Listeria innocua NCTC 11288. In artificially contaminated ready-to-eat pork ham, BLIS was effective in inhibiting the growth of L. seeligeri NCTC 11289 for 6 days (counts from 1.74 to 0.00 log CFU/g) and ensured lower weight loss (2.7%) and lipid peroxidation (0.63 mg MDA/kg) of samples compared with the control (3.0%; 1.25 mg MDA/kg). At the same time, coloration of ham samples in terms of luminosity, redness, and yellowness as well as discoloration throughout cold storage was not influenced by BLIS or Nisaplin taken as a control. These results suggest the potential use of P. pentosaceus BLIS as a biopreservative in meat and other food processing industries

Optimized nickase- and nuclease-based prime editing in human and mouse cells

Precise genomic modification using prime editing (PE) holds enormous potential for research and clinical applications. In this study, we generated all-inone prime editing (PEA1) constructs that carry all the components required for PE, along with a selection marker. We tested these constructs (with selection) in HEK293T, K562, HeLa and mouse embryonic stem (ES) cells. We discovered that PE efficiency in HEK293T cells was much higher than previously observed, reaching up to 95% (mean 67%). The efficiency in K562 and HeLa cells, however, remained low. To improve PE efficiency in K562 and HeLa, we generated a nuclease prime editor and tested this system in these cell lines as well as mouse ES cells. PE-nuclease greatly increased prime editing initiation, however, installation of the intended edits was often accompanied by extra insertions derived from the repair template. Finally, we show that zygotic injection of the nuclease prime editor can generate correct modifications in mouse fetuses with up to 100% efficiency.Fatwa Adikusuma, Caleb Lushington, Jayshen Arudkumar, Gelshan I. Godahewa, Yu C.J. Chey, Luke Gierus, Sandra Piltz, Ashleigh Geiger, Yatish Jain, Daniel Reti, Laurence O.W. Wilson, Denis C. Bauer, and Paul Q. Thoma

Antibacterial and antifungal activity of crude and freeze-dried bacteriocin-like inhibitory substance produced by Pediococcus pentosaceus

Pediococcus pentosaceus LBM 18 has shown potential as producer of an antibacterial and antifungal bacteriocin-like inhibitory substance (BLIS). BLIS inhibited the growth of spoilage bacteria belonging to Lactobacillus, Enterococcus and Listeria genera with higher activity than Nisaplin used as control. It gave rise to inhibition halos with diameters from 9.70 to 20.00 mm, with Lactobacillus sakei being the most sensitive strain (13.50\u201320.00 mm). It also effectively suppressed the growth of fungi isolated from corn grain silage for up to 25 days and impaired morphology of colonies by likely affecting fungal membranes. These results point out that P. pentosaceus BLIS may be used as a new promising alternative to conventional antibacterial and antifungal substances, with potential applications in agriculture and food industry as a natural bio-controlling agent. Moreover, cytotoxicity and cell death induction tests demonstrated cytotoxicity and toxicity of BLIS to human colon adenocarcinoma Caco-2cells but not to peripheral blood mononuclear cells, with suggests possible applications of BLIS also in medical-pharmaceutical applications