Recent advances in the study of the CRISPR/Cas9 system have provided a precise and versatile approach for genome editing in various species. However, the applicability and efficiency of this method in large animal models, such as the goat, have not been extensively studied. Here, by co-injection of one-cell stage embryos with Cas9 mRNA and sgRNAs targeting two functional genes (MSTN and FGF5), we successfully produced gene-modified goats with either one or both genes disrupted. The targeting efficiency of MSTN and FGF5 in cultured primary fibroblasts was as high as 60%, while the efficiency of disrupting MSTN and FGF5 in 98 tested animals was 15% and 21% respectively, and 10% for double gene modifications. The on- and off-target mutations of the target genes in fibroblasts, as well as in somatic tissues and testis of founder and dead animals, were carefully analyzed. The results showed that simultaneous editing of several sites was achieved in large animals, demonstrating that the CRISPR/Cas9 system has the potential to become a robust and efficient gene engineering tool in farm animals, and therefore will be critically important and applicable for breeding.
Mechanical and barrier properties of corn zein films with butylated hydroxyanisole (BHA), bacterial enzyme, and emulsifier additives were investigated and compared to those of polyvinylidine chloride (PVDC) films. No significant differences (P Ͻ 0.05) occurred in elongation, tensile strength, or Young's modulus among corn zein films. PVDC had the lowest apparent water vapor permeability (WVP app ), and no significant differences in WVP app were found between zein films and PVDC. Cooked turkey breast was packaged, evaluated by a trained sensory panel, and analyzed for hexanal content. Turkey breast wrapped in corn zein film with antioxidant and emulsifier had lower hexanal content than samples packaged in PVDC.
Three semicontinuous continuous stirred-tank reactors (CSTR) operating at mesophilic conditions (35°C) were used to investigate the effect of hydraulic retention time (HRT) on anaerobic digestion of wheat straw. The results showed that the average biogas production with HRT of 20, 40, and 60 days was 46.8, 79.9, and 89.1 mL/g total solid as well as 55.2, 94.3, and 105.2 mL/g volatile solids, respectively. The methane content with HRT of 20 days, from 14.2% to 28.5%, was the lowest among the three reactors. The pH values with HRT of 40 and 60 days were in the acceptable range compared to that with HRT of 20 days. The propionate was dominant in the reactor with HRT of 20 days, inhibiting the activities of methanogens and causing the lower methane content in biogas. The degradation of cellulose, hemicellulose, and crystalline cellulose based on XRD was also strongly influenced by HRTs.
Raw yolk of 1-d-old eggs was either subjected to linear electron beam irradiation at approximately 2.5 kGy dosage or not processed. Both irradiated and nonprocessed egg yolk samples were stored at -15 C after irradiation. Testing was conducted on 0, 1, 7, 15, 30, and 60 d of storage. Development of storage modulus (G') was delayed in irradiated samples after 7 d, which suggests that less structure was developed in irradiated egg yolk than in nonprocessed egg yolk during storage. Irradiated samples retained more soluble protein within the first 7 d and showed slightly improved emulsion capacity over that from nonprocessed samples. However, irradiated egg yolk was less bright than nonprocessed samples. No differences were observed in SDS-PAGE patterns of soluble proteins and delipidized low density lipoprotein (LDL). The LDL isolated from irradiated liquid egg yolk showed no difference in N-terminal amino acids compared to that of nonprocessed egg yolk, indicating no detectable cleavage of LDL. However, the denaturation temperature of irradiated samples at Day 0 shifted about 1 C lower than that of the nonprocessed sample. Results indicated that electron beam irradiation did not cause significant physical, chemical or functional changes of egg yolk, or cleavage of egg yolk protein. Therefore, electron beam irradiation could serve as a preservation method for liquid egg yolk.
The aim of this study was to investigate the influence of lipoxygenase-less (LOX-less) barley malt on the quality of wort and beer, with the main focus on beer flavour stability. In the current study, pilot-scale (1000 L) brewing trials were conducted with a control barley malt AC Metcalfe and a LOX-less barley malt, PolarStar. The results clearly indicated that the LOX-less barley malt showed less nonenal potential than the control, although LOX activities in both barley malts were relatively low. The beer brewed from the LOX-less barley malt contained much lower concentrations of trans-2-nonenal (T2N) and gamma-nonalactone, especially after the (forced or natural) aging of the beer, compared with the beer brewed under the same conditions using the control malt. The sensory panel evaluation indicated similar results in the general flavour profile. The freshness scores of beer brewed from the LOX-less malt were higher than those from the control malt, and this was more pronounced after forced aging. In addition, the beer brewed from LOX-less malt had a much better foam stability, almost 30 s (NIBEM test). These results confirm that the use of the LOX-less barley malt was beneficial to beer flavour stability and foam stability.
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