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The conventional commercial hatcheries used today do not allow the newly hatched chicks to consume feed or water. Combined with natural variation in hatching time, this can lead to early hatched chicks being feed-deprived for up to 72 h before being unloaded at the rearing site. This study investigated the effects of hatching time on time to first feed intake and development of organs, digestive enzymes and productivity in terms of growth and feed conversion ratio in chicks hatched on-farm. Chicks were divided into three hatching groups (early, mid-term and late), and assessed over a full production cycle of 34 days. The results revealed that chicks remain inactive for a considerable amount of time before engaging in eating-related activities. Eating activity of 5% (i.e. when 5% of birds in each hatching group were eating or standing close to the feeder) was recorded at an average biological age (BA) of 25.4 h and a proportion of 50% birds with full crop was reached at an average BA of 30.6 h. Considering thatween hatching groups, this study showed that the hatchlings seemed capable of compensating for these as they grew.Recent studies have shown that chromium (Cr) could alleviate the negative effects of heat stress on livestock and poultry, but there is little information available to laying ducks. This study aimed to investigate the effects of dietary addition of chromium propionate on laying performance, egg quality, serum biochemical parameters and antioxidant status of laying ducks under hot (average 32 °C) and humid (average 75% relative humidity) summer conditions. ML265 concentration A total of 900 66-week-old weight- and laying-matched Shanma laying ducks were randomly divided into five treatments, each with 6 replicates of 30 individually caged birds. The birds were fed either a basal diet or the basal diet supplemented with either 200, 400, 600, or 800 μg/kg Cr as chromium propionate. All laying ducks were given feed and water ad libitum for 5 weeks. The results showed that dietary supplementation with chromium propionate significantly increased the laying rate and yolk colour score (P  less then  0.05). Treatment with 400 μg/kg Cr as μg/kg could beneficially affect the laying rate, egg quality and antioxidant function, as well as modulate the blood biochemical parameters of laying ducks under heat stress conditions.Electron microscopy has consistently played an important role in the description of pore-forming protein systems. The discovery of pore-forming proteins has depended on visualization of the structural pores formed by their oligomeric protein complexes, and as electron microscopy has advanced technologically so has the degree of insight it has been able to give. This review considers a large number of published studies of pore-forming complexes in prepore and pore states determined using single-particle cryo-electron microscopy. Sample isolation and preparation, imaging and image analysis, structure determination and optimization of results are all discussed alongside challenges which pore-forming proteins particularly present. The review also considers the use made of cryo-electron tomography to study pores within their membrane environment and which will prove an increasingly important approach for the future.In vivo proteins fold mainly as they emerge from the ribosome or as they emerge from a membrane translocon. Membrane translocation in particular poses technical challenges to the study of the associated protein folding processes. Recently we have developed a single-molecule methodology that allows the capture of a single protein molecule through a membrane translocon with biotinylated oligonucleotides covalently bound at its N- and C- terminus using streptavidin. The resulting rotaxane can be driven forwards and backwards changing the voltage polarity, and carefully planned experiments allow inference of the folding pathway. Here we will discuss the details of a simplified methodological approach.We report here on the nanopore resistive pulse sensing (Np-RPS) method, involving pore-forming toxins as tools for polymer analytics at single molecule level. Np-RPS is an electrical method for the label-free detection of single molecules. A molecule interacting with the pore causes a change of the electrical resistance of the pore, called a resistive pulse, associated with a measurable transient current blockade. The features of the blockades, in particular their depth and duration, contain information on the molecular properties of the analyte. We first revisit the history of Np-RPS, then we discuss the effect of the configuration of the molecule/nanopore interaction on the molecular information that can be extracted from the signal, illustrated in two different regimes that either favor molecular sequencing or molecular sizing. Specifically, we focus on the sizing regime and on the use of two different pore-forming toxins, staphylococcal α-hemolysin (αHL) and aerolysin (AeL) nanopores, for the characterization of water-soluble polymers (poly-(ethylene glycol), (PEG)), homopeptides, and heteropeptides. We discuss how nanopore sizing of polymers could be envisioned as a new approach for peptide/protein sequencing.Pore-forming toxins are used in a variety of biotechnological applications. Typically, individual membrane proteins are reconstituted in artificial lipid bilayers where they form water-filled nanoscale apertures (nanopores). When a voltage is applied, the ionic current passing through a nanopore can be used for example to sequence biopolymers, identify molecules, or to study chemical or enzymatic reactions at the single-molecule level. Here we present strategies for studying individual enzymes and measuring molecules, also in highly complex biological samples such as blood.Cholesterol is a major component of the plasma membranes (PMs) of animal cells, comprising 35-40mol% of total PM lipids. Recent studies using cholesterol-binding bacterial toxins such as domain 4 of Anthrolysin O (ALOD4) and fungal toxins such as Ostreolysin A (OlyA) have revealed new insights into the organization of PM cholesterol. These studies have defined three distinct pools of PM cholesterol-a fixed pool that is essential for membrane integrity, a sphingomyelin (SM)-sequestered pool that can be detected by OlyA, and a third pool that is accessible and can be detected by ALOD4. Accessible cholesterol is available to interact with proteins and transport to the endoplasmic reticulum (ER), and controls many cellular signaling processes including cholesterol homeostasis, Hedgehog signaling, and bacterial and viral infection. Here, we provide detailed descriptions for the use of ALOD4 and OlyA, both of which are soluble and non-lytic proteins, to study cholesterol organization in the PMs of animal cells. Furthermore, we describe two new versions of ALOD4 that we have developed to increase the versatility of this probe in cellular studies.