This paper describes, in detail, the development of a novel, low-cost, and flexible drift tube (DT) along with an associated ion mobility spectrometer system. The DT is constructed from a flexible ...printed circuit board (PCB), with a bespoke “dog-leg” track design, that can be rolled up for ease of assembly. This approach incorporates a shielding layer, as part of the flexible PCB design, and represents the minimum dimensional footprint conceivable for a DT. The low thermal mass of the polyimide substrate and overlapping electrodes, as afforded by the dog-leg design, allow for efficient heat management and high field linearity within the tube–achieved from a single PCB. This is further enhanced by a novel double-glazing configuration which provides a simple and effective means for gas management, minimizing thermal variation within the assembly. Herein, we provide a full experimental characterization of the flexible DT ion mobility spectrometer (Flex-DT-IMS) with corresponding electrodynamic (Simion 8.1) and fluid dynamic (SolidWorks) simulations. The Flex-DT-IMS is shown to have a resolution >80 and a detection limit of low nanograms for the analysis of common explosives (RDX, PETN, HMX, and TNT).
Although environmental enrichment is known to improve laboratory rodent wellbeing and enhance scientific data collection, relatively little is known with regards to the type of enrichment that might ...be useful for zebrafish (Danio rerio). Therefore, this study explored if zebrafish displayed preferences for a range of enrichments, including substrates, artificial plants, combinations thereof and airstones. Tanks divided into two compartments containing different enrichment cues were used to determine the preferences of zebrafish housed in pairs and groups of eight. When comparing time spent in enriched versus barren compartments, dominant individuals in a pair displayed a preference for substrate and behaviourally excluded the subordinate (p < 0.05). In groups there was a preference for all substrate (p < 0.01) and plant (p < 0.05) enrichments over barren conditions. The strongest preference was for gravel substrate and images of gravel attached to the bottom of the tank. When preferences were compared for different enrichments, gravel (both sexes, p < 0.01) again emerged as the cue attracting the most significant preferences, with any combination featuring gravel substrate preferred over any combination featuring sand (p < 0.05). The study has demonstrated that zebrafish reared in barren conditions preferred structural enrichment over standard conditions; however, when fish were held in pairs this was influenced by dominance status and in groups this was influenced by gender.
In rapidly growing animals there are numerous selective pressures and developmental constraints underpinning the ontogenetic development of muscle-tendon morphology and mechanical properties. Muscle ...force generating capacity, tendon stiffness, elastic energy storage capacity and efficiency were calculated from muscle and tendon morphological parameters and
tendon mechanical properties obtained from a growth series of ostrich cadavers. Ontogenetic scaling relationships were established using reduced major axis regression analysis. Ostrich pelvic limb muscle mass and cross-sectional area broadly scaled with positive allometry, indicating maintained or relatively greater capacity for maximum isometric force generation in larger animals. The length of distal limb tendons was found to scale with positive allometry in several tendons associated with antigravity support and elastic energy storage during locomotion. Distal limb tendon stiffness scaled with negative allometry with respect to body mass, with tendons being relatively more compliant in larger birds. Tendon material properties also appeared to be size-dependent, suggesting that the relative increased compliance of tendons in larger ostriches is due in part to compensatory distortions in tendon material properties during maturation and development, not simply from ontogenetic changes in tendon geometry. Our results suggest that the previously reported increase in locomotor economy through ontogeny in the ostrich is due to greater potential for elastic energy storage with increasing body size. In fact, the rate of this increase may be somewhat greater than the conservative predictions of previous studies, thus illustrating the biological importance of elastic tendon structures in adult ostriches.
The state of an animal prior to the application of a noxious stimulus can have a profound effect on their nociceptive threshold and subsequent behaviour. In mammals, the presence of acute stress ...preceding a painful event can have an analgesic effect whereas the presence of chronic stress can result in hyperalgesia. While considerable research has been conducted on the ability of stress to modulate mammalian responses to pain, relatively little is known about fish. This is of particular concern given that zebrafish (
) are an extensively used model organism subject to a wide array of invasive procedures where the level of stress prior to experimentation could pose a major confounding factor. This study, therefore, investigated the impact of both acute and chronic stress on the behaviour of zebrafish subjected to a potentially painful laboratory procedure, the fin clip. In stress-free individuals, those subjected to the fin clip spent more time in the bottom of the tank, had reduced swimming speeds and less complex swimming trajectories; however, these behavioural changes were absent in fin-clipped fish that were first subject to either chronic or acute stress, suggesting the possibility of stress-induced analgesia (SIA). To test this, the opioid antagonist naloxone was administered to fish prior to the application of both the stress and fin-clip procedure. After naloxone, acutely stressed fin-clipped zebrafish exhibited the same behaviours as stress-free fin-clipped fish. This indicates the presence of SIA and the importance of opioid signalling in this mechanism. As stress reduced nociceptive responses in zebrafish, this demonstrates the potential for an endogenous analgesic system akin to the mammalian system. Future studies should delineate the neurobiological basis of stress-induced analgesia in fish.
Paper-based microfluidic channels were created from solid wax printing, and the resultant 2D wax-printed paper substrates were used for paper spray (PS) mass spectrometry (MS) analysis of small ...organic compounds. Controlling fluid flow at the tip of the wax-printed paper triangles enabled the use of lower spray voltages (0.5-1 kV) and extended signal lifetime (10 minutes) in PS-MS. High sensitivity (sub ng mL(-1) levels) and quantitation precision (<10% RSD) have been achieved in the analysis of illicit drugs in 4 μL of raw urine (fresh and dry), as well as corrosion inhibitors and pesticides in water samples. The reported study encourages the future development of disposable 3D microfluidic paper-based analytical devices, which function with simple operation but capable of on-chip analyte detection by MS; such a device can replace the traditional complex laboratory procedures for MS analysis to enable on-site in situ sampling with portable mass spectrometers.
Fish are used in a variety of experimental contexts often in high numbers. To maintain their welfare and ensure valid results during invasive procedures it is vital that we can detect subtle changes ...in behaviour that may allow us to intervene to provide pain-relief. Therefore, an automated method, the Fish Behaviour Index (FBI), was devised and used for testing the impact of laboratory procedures and efficacy of analgesic drugs in the model species, the zebrafish. Cameras with tracking software were used to visually track and quantify female zebrafish behaviour in real time after a number of laboratory procedures including fin clipping, PIT tagging, and nociceptor excitation via injection of acetic acid subcutaneously. The FBI was derived from activity and distance swum measured before and after these procedures compared with control and sham groups. Further, the efficacy of a range of drugs with analgesic properties to identify efficacy of these agents was explored. Lidocaine (5 mg/L), flunixin (8 mg/L) and morphine (48 mg/L) prevented the associated reduction in activity and distance swum after fin clipping. From an ethical perspective, the FBI represents a significant refinement in the use of zebrafish and could be adopted across a wide range of biological disciplines.
Fish production is increasingly important to global food security. A major factor in maintaining health, productivity and welfare of farmed fish is the establishment and promotion of a stable and ...beneficial intestinal microbiota. Understanding the effects of factors such as host and environment on gut microbial community structure is essential for developing strategies for stimulating the establishment of a health-promoting gut-microbiota. We compared intestinal microbiota of common carp and rainbow trout, two fish with different dietary habits, sourced from various farm locations. There were distinct differences in the gut microbiota of carp and trout intestine. The microbiota of carp was dominated by Fusobacteriia and Gammaproteobacteria, while the trout microbiota consisted predominantly of Mollicutes and Betaproteobacteria. The majority of bacterial sequences clustered into a relatively low number of operational taxonomic units (OTUs) revealing a comparatively simple microbiota, with Cetobacterium, Aeromonas and Mycoplasma being highly abundant. Within each species, fish from different facilities were found to have markedly similar predominant bacterial populations despite distinctly different rearing environments, demonstrating intra-species uniformity and significant influence of host selectivity. This study demonstrates that in fish the host species imparts substantial impact in shaping the community structure of the intestinal microbiota.
Peak power demand strains electrical grids and increases cost of electricity generation, transmission and distribution infrastructure. Many studies have examined ways of reducing peak power demand, ...including modification of room air temperature setpoints or reduction of lighting levels. However, very few or no studies have examined the peak power reduction resource offered by temporary curtailment of building ventilation systems. For this reason, we conducted a simulation campaign in which we examined the resource offered by temporary ventilation curtailment in commercial buildings of different use types across the United States and in residences in the state of California, with the essential constraint that any changes resulted in air quality acceptable to occupants through additional ventilation to compensate for the curtailment. To do this, we employed previously validated building models implemented in the airflow and contaminant transport tool CONTAM and building thermal and systems modeling tool EnergyPlus, in some cases co-simulated. Results show savings are highly dependent on building type and climate but range from 0–2 W/ft2 and up to 40% of total peak building power demand. Depending on building type, this power shed can be conducted for 1.5–8 h before acute exposure or odor concerns are expected, assuming a safety factor of 2. This reduction is of the same magnitude as that offered by thermal control strategies such as setpoint increase, or from lighting reduction strategies.
A novel strategy for the direct analysis of non-conjugated steroids in water using paper spray mass spectrometry (PS-MS) has been developed. PS-MS was used in the identification and quantification of ...non-conjugated (free) steroids in fish tank water samples. Data shown herein indicates that individual amounts of free steroids can be detected in aqua as low as; 0.17 ng/µL, 0.039 ng/µL, 0.43 ng/µL, 0.0076 ng/µL for aldosterone, corticosterone, cortisol, and β-estrone, respectively, and with an average relative standard deviation of ca. < 10% in the positive ion mode using PS-MS/MS. Direct detection of free steroids in a raw water mixture, from aquaculture, without prior sample preparation is demonstrated. The presence of free steroids released in fish water samples was confirmed via tandem mass spectrometry using collision-induced dissociation. This approach shows promise for rapid and direct water quality monitoring to provide a holistic assessment of non-conjugated steroids in aqua.
Stressful experiences can have detrimental effects on many aspects of health and wellbeing. The zebrafish (Danio rerio) is a widely used model for stress research and a stress phenotype can be ...induced by manipulating the environmental conditions and social interactions. In this study we have combined a zebrafish stress model with the measurement of degradation rates of soluble cardiac muscle proteins. The results showed that the greater the stress response in the zebrafish the lower the level of overall protein degradation. On comparing the rates of degradation for individual proteins it was found that four main pathways were altered in response to stress conditions with decreased degradation for proteins involved in glucose metabolism, gluconeogenesis, the ubiquitin-proteasome system (UPS) and peroxisomal proliferator-activated receptor (PPAR) signalling pathways. Taken together, these data indicate that under stress conditions zebrafish preserve cardiac muscle proteins required for the ‘fight or flight’ response together with proteins that play a role in stress mitigation.
This study is the first to investigate the impact of stressful experiences on the dynamics of protein turnover in cardiac muscle. Using an established zebrafish model of human stress it has been possible to map key pathways at the protein level. The results show that the rates of degradation of cardiac proteins involved in glucose metabolism, UPS activity, hypoxia and PPAR signalling are decreased in stressed zebrafish. These findings indicate that proteins involved in the ‘fight or flight’ response to stress are conserved by the heart together with proteins that play a role in stress mitigation. This work provides the basis for more detailed investigations aimed at understanding the molecular effects of stress, which has implications for human health and disease.
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•The degradation rates of cardiac proteins measured in a zebrafish model of stress.•Decrease in individual protein degradation during stressed phenotype•Proteins involved in glucose metabolism and ubiquitin-proteasome system•Indicates preservation of cardiac proteins for the ‘fight or flight’ response
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