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.     

Composition and fatty acid profile of milk from cows supplemented with pressed oilseed cake

This study compared the productive and nutritional parameters of milk from crossbred lactating cows managed on Panicum maximum Jacq. cv. Tanzania and with a diet supplemented with different pressed oilseed cakes. The supplements used were as follows: peanut cake, sunflower cake and palm kernel cake for replacement of soybean meal. Sixteen cows with an average weight of 544 ± 57 kg and producing 8 ± 1.4 L of milk per day were used in this study. The animals were randomly assigned to the treatments according to a Latin square design repeated over time, with four treatments, 16 animals and four experimental periods. Supplementation of the diet with peanut cake, sunflower cake and palm kernel cake compared with soybean meal in the diet of cows did not affect the average daily production or composition of the milk. The palm kernel cake promoted an increase in lauric fatty acids (C_12:0) and palmitoleic acids (C_16:1) (5.02 and 1.65%, respectively) compared with peanut cake and sunflower cake (4.13 and 4.01%, respectively). The levels of oleic fatty acids (C_18:1) were higher for the sunflower cake and palm kernel cake supplements (26.01 and 25.01%, respectively) compared with peanut cake (23.11%). The replacement of soybean meal with sunflower cake and palm kernel cake improved the nutritional quality of the milk, with lower concentrations of saturated fatty acids and higher concentrations of unsaturated fatty acids, without compromising the production or nutritional composition of the milk. © 2015 Japanese Society of Animal Science     

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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Sodicity and salinity in a Brazilian Oxisol cultivated with sugarcane irrigated with wastewater

Changes in soil sodicity-salinity parameters are one of the most characteristic alterations after treated sewage effluent (TSE) irrigation in agro-systems. Considering the importance of these parameters for agricultural management, as well as the economical value of sugarcane for Brazil, the present study aimed at evaluating effects on soil sodicity and salinity under tropical conditions over 16 months of TSE irrigation in a sugarcane plantation at Lins, São Paulo State, Brazil. Soil samplings were carried out in February 2005 (before planting), December 2005 (after 8 months of TSE irrigation) and September 2006 (after 16 months of TSE irrigation) following a complete block design with four treatments and four replicates. Treatments consisted of: (i) control, without TSE irrigation; (ii) T100, T150 and T200, with TSE irrigation supplying 100% (0% surplus, total of 2524 mm), 150% (50% surplus, total of 3832 mm) and 200% (100% surplus, total of 5092 mm) of crop water demand, respectively. Compared to initial soil conditions, at the end of the experiment increases of exchangeable sodium (from 2.4 to 5.9 mmol_c kg⁻¹), exchangeable sodium percentage (ESP) (from 8 to 18%), soluble Na (from 1.4 to 4.7 mmol L⁻¹) and sodium adsorption ratio (SAR) of soil solution (from 3.6 to 12.6 (mmol L⁻¹)^0.5) were found in the soil profile (0-100 cm) as an average for the irrigated plots due to high SAR of TSE. Associated with the increments were mostly significant increases in clay dispersion rates at depths 0-10, 10-20 and 20-40 cm. Electrical conductivity (EC) of soil solution increased during the TSE irrigation period whereas at the end of the experiment, after short term discontinuation of irrigation and harvest, EC in the topsoil (0-10 and 10-20 cm) decreased compared to the previous samplings. Moreover, despite increasing sodicity over time mainly insignificant differences within the different irrigated treatments were found in December 2005 and September 2006. This suggests that independent of varying irrigation amounts the increasing soil sodicity over time were rather caused by the continuous use of TSE than by its quantity applied. Moreover, also plant productivity showed no significant differences within the TSE irrigated plots. The study indicates that monitoring as well as remediation of soil after TSE irrigation is required for a sustainable TSE use in order to maintain agricultural quality parameters.