The effects of supplemental graded levels of neutral phytase using pretreatment and spraying methods on the apparent digestibility coefficients (ADCs) of protein, minerals and amino acids were investigated in grass carp (Ctenopharyngodon idellus) (mean weight, 20.92 ± 1.04 g). Seven experimental diets were prepared as: no‐phytase supplementation (control), pretreatment with phytase dose of 500 U (PP500), 1000 U (PP1000) and 1500 U (PP1500) per kg diet. With spraying method, the phytase was added to feed at the same three grades, namely 500 U (PS500), 1000 U (PS1000) and 1500 U (PS1500) per kg diet. After an 8‐week feeding trial, addition of phytase improved the crude protein, crude lipid, dry matter, ash, phosphorus and calcium digestibilities in comparison with the control, especially in the diets where phytase addition level was above 1000 U kg−1 (P < 0.05). Phytase supplementation, both the pretreatment and spraying, significantly enhanced the ADCs of essential amino acid (EAA) and non‐essential amino acid. The ADCs of the nutrients were significantly influenced by phytase supplementation methods, supplementation levels and interaction of methods and levels. Based on the results, supplemental dietary phytase, regardless of application methodology, improved the ADCs of nutrients in grass carp, and the pretreatment mode of phytase application was more effective than spraying at the same phytase level. We suggest that the optimal level of phytase pretreatment is 1000 U kg−1 diet, and the best addition level of phytase spraying is 1500 U kg−1 diet or more phytase is required for spraying.
The existence of membrane contact sites (MCS) has been reported in different systems in the past decade, and their importance has been recognised by the cell biology community. Amongst all endomembrane structures, the endoplasmic reticulum (ER) plays vital roles in organising the organelle interaction network with the plasma membrane (PM), Golgi bodies, mitochondria, plastids, endosomes and autophagosomes. A number of methods have been used to study the establishment and functions of these interactions, among them, light microscopy appears to be one of the most effective approaches. Here, we present an overview of the discovery of ER-PM contact sites, and highlight the latest developments in light microscopical-based techniques that can be used for their study.
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