Optimizing injection time of GFP plasmid into sheep zygote

Microinjection of exogenous DNA into the cytoplasm of matured oocytes or zygotes is a promising technique to generate transgenic animals. However, the data about the microinjection time and procedure in sheep are limited and have not treated in detail. To obtain more in-depth information, the Sarda sheep oocytes from abattoir-derived ovaries were subjected to IVM and IVF. Then, the GFP plasmid as a reporter gene was injected into the cytoplasm of MII oocytes (n: 95) and zygotes at different post-insemination intervals (6–8 hpi, n: 120; 8–10 hpi, n: 122; 10–12 hpi, n: 110 and 12–14 hpi, n: 96). There were no significant differences in the cleavage rates between the groups. However, blastocyst rate of injected zygotes at all-time intervals was significantly lower than injected MII oocytes and control group (p < 0.05). Interestingly, the proportion of GFP-positive embryos was higher at 8–10 hpi compared with other injected groups (4 % versus 0 %, p  < 0.01). Among these, the proportion of mosaic embryos was high and two of those embryos developed to the blastocyst stage. In conclusion, we settled on the cytoplasmic microinjection of GFP plasmid at 8–10 hpi as an optimized time point for the production of transgenic sheep and subsequent experiments.     

The Production of Microbiologically Safe Effluents for Wastewater Reuse in the Middle East and North Africa

Wastewater treatment in waste stabilization ponds (WSP) is a very efficient, low cost, low maintenance process which is well able to produce an effluent quality that meets the World Health Organization's recommendations for wastewater reuse for crop irrigation. Treatment in anaerobic and facultative ponds is required for restricted irrigation, with further treatment in maturation ponds for unrestricted irrigation. However, it is shown that the land requirements for the latter are at least twice that for the former. Unrestricted irrigation should, therefore, only be selected if it is economically viable. The use of wastewater storage and treatment reservoirs (WSTR), after pretreatment in anaerobic ponds, is advantageous in that it permits the whole year's wastewater to be used for irrigation, so allowing a much greater quantity of crops to be produced. A hybrid WSP-WSTR system can be used which produces effluents safe for both restricted and unrestricted irrigation.     

Ultrasound-Assisted Extraction for the Recovery of Carotenoids from Guava’s Pulp and Waste Powders

Plant Foods for Human Nutrition - In this work, lycopene- and β-carotene-rich extracts were obtained from guava’s pulp and waste powders using maceration (ME), ultrasonic bath (BUAE,...     

Mesohabitat associations of the threatened San Marcos salamander (Eurycea nana) across its geographic range

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