Automated zebrafish chorion removal and single embryo placement: optimizing throughput of zebrafish developmental toxicity screens

The potential of the developing zebrafish model for toxicology and drug discovery is limited by inefficient approaches to manipulating and chemically exposing zebrafish embryos-namely, manual placement of embryos into 96- or 384-well plates and exposure of embryos while still in the chorion, a barrier of poorly characterized permeability enclosing the developing embryo. We report the automated dechorionation of 1600 embryos at once at 4 h postfertilization (hpf) and placement of the dechorionated embryos into 96-well plates for exposure by 6 hpf. The process removed ≥95% of the embryos from their chorions with 2% embryo mortality by 24 hpf, and 2% of the embryos malformed at 120 hpf. The robotic embryo placement allocated 6-hpf embryos to 94.7% ± 4.2% of the wells in multiple 96-well trials. The rate of embryo mortality was 2.8% (43 of 1536) from robotic handling, the rate of missed wells was 1.2% (18 of 1536), and the frequency of multipicks was <0.1%. Embryo malformations observed at 24 hpf occurred nearly twice as frequently from robotic handling (16 of 864; 1.9%) as from manual pipetting (9 of 864; 1%). There was no statistical difference between the success of performing the embryo placement robotically or manually.     

Limited evidence of zebra mussel (Dreissena polymorpha) consumption by freshwater drum (Aplodinotus grunniens) in Lake Winnipeg

Freshwater drum (Aplodinotus grunniens) may be a predator of the invasive zebra mussel (Dreissena polymorpha), which established in Lake Winnipeg in 2013. In this study, the diets, trophic position, and growth of 51 freshwater drum collected in 2019 (six years post-zebra mussel invasion) were compared to 64 freshwater drum sampled in 2000. Benthic insect larvae were the dominant food items in both years. Although mollusks occur in high densities in Lake Winnipeg, they were only consumed by a few freshwater drum in either sample year. Zebra mussels were not a frequent prey item in 2019 as they were only consumed by four of the sampled freshwater drum. Stable isotope analysis of white muscle tissue yielded similar δ¹³C and δ¹⁵N values in both years and were consistent with a benthic, insectivorous diet. Length-at-age data derived from otoliths revealed that the 2019 population had at least an equal growth rate to the 2000 population. Weight-at-length data suggested that fish condition was greater in 2019 than in 2000, which coincided with increased benthic macroinvertebrate density in Lake Winnipeg. Based on these findings, Lake Winnipeg freshwater drum continue to feed predominantly on insect larvae and not zebra mussels.     

Modeling heavy metal mass releases from urban battery litter

Consumer batteries littered on urban pavements release metals of environmental significance (Ag, Cd, Cr, Cu, Hg, Li, Mn, Ni, Pd, Ti, Zn) to stormwater runoff. Predicting the mass loading of any one metal is difficult because of the random composition of battery litter. However, when littering is modeled as a conditional filtered Poisson process, bounds may be estimated for the mean and variance of site mass loading for any metal if the site litter rate and battery product contributions are known. Site-specific data on the battery brand distribution in litter can improve load estimates, but statistics computed from 5500 littered batteries collected in the Cleveland area may be used to approximate the brand distribution. Zinc load calculations based on battery litter size, type and brand discretizations are implemented in a model titled BLML and illustrated for a case-study location. Results indicate that, at some urban sites, zinc released from battery litter can be the largest source of zinc in urban pavement runoff.     

Synthesis and applications of molybdenum (IV) oxide

Molybdenum dioxide (MoO₂) is a transition metal oxide with unusual metal-like electrical conductivity and high catalytic activity toward reforming hydrocarbons. This review covers the synthesis techniques used to fabricate MoO₂ in a variety of morphologies and particle sizes. Processing from molybdenum ore and reduction from MoO₃ are also covered, with emphasis on reduction mechanisms and kinetic considerations. Discussions of various solution-based and gas phase synthesis techniques shed light on strategies to achieve various unique morphologies, which leads into a brief discussion of nanoscale MoO₂. Nanoscale MoO₂ is of interest for important technological applications including catalysts for partial oxidation of hydrocarbons, solid oxide fuel cell anodes, and high-capacity reversible lithium ion battery anodes.     

Trout‐Skin Gelatin‐Based Edible Films Containing Phenolic Antioxidants: Effect on Physical Properties and Oxidative Stability of Cod‐Liver Oil Model Food

Abstract: Trout‐skin (Oncorhynchus mykiss) gelatin‐based films containing antioxidants (epigallocatechin gallate (EGCG), 50 and 250 ppm w/w) and green tea powder (1% and 20% w/w of gelatin) were tested for tensile strength, elastic modulus, and elongation, and oxygen and water vapor transmission rates, in vitro antioxidant activity using the DPPH (2,2‐diphenyl‐1‐picrylhydrazyl) assay and effect on stabilizing cod‐liver oil held under mild thermal abuse conditions. Cod‐liver oil overlaid with films was stored at 40 °C for 20 d and analyzed for peroxide value (PV) and thiobarbituric acid reactive substances (TBARS). Antioxidant activity was retained in films containing green tea powder, but was reduced (P < 0.05) in EGCG films (20 d, 23 °C). Water vapor transmission rate of the films incorporated with antioxidants did not change significantly (P > 0.05), but the oxygen transmission rate for films with 50 ppm EGCG and 20% green tea powder was significant (P < 0.05). Other physical properties varied with antioxidant incorporation. The TBARS and PV of control oil increased from 0.05 ± 0.01 to 4.71 ± 0.30 g MDA/kg oil and from 3.6 ± 0.2 to 178.3 ± 24.5 millieq peroxides/kg oil, respectively, after 20 d. For cod‐liver oil covered with control or antioxidant‐containing films, TBARS remained below 0.37 g MDA/kg oil and PV below 7 millieq peroxides/kg oil. Incorporation of antioxidants to the films did not reduce oil oxidation (P > 0.05) at the levels tested and this was confirmed by activation energy calculations. The rate of oil oxidation was more dependent upon the inherent oxygen barrier property of the films than the presence of antioxidants. Practical Application: This research has the potential to enhance the utilization of fish skins, a valuable food processing by‐product, as edible films with natural antioxidants to extend the shelf life of foods. The film physical properties and barrier to oxygen and water are investigated.     

The House Mouse (Mus musculus L.) Exerts Strong Differential Grain Consumption Preferences among Hard Red and White Spring Wheat (Triticum aestivum L.) Varieties in a Single‐Elimination Tournament Design

Wheat (Triticum aestivum L.) plays a central role in the health and nutrition of humans. Yet, little is known about possible flavor differences among different varieties. We have developed a model system using the house mouse (Mus musculus L.) to determine feeding preferences as a prelude to extending results to human sensory analysis. Here, we examine the application of a single‐elimination tournament design to the analysis of consumption preferences of a set of hard red and hard white spring wheat varieties. A single‐elimination tournament design in this case pairs 2 wheat varieties and only 1 of the 2 is advanced to further tests. Preferred varieties were advanced until an overall “winner” was identified; conversely, less desirable varieties were advanced such that an overall “loser” was identified. Hollis and IDO702 were the winner and loser, respectively, for the hard red varieties, and Clear White 515 and WA8123 were the winner and loser, respectively, for the hard white varieties. When using the more powerful protocol of 14 mice and a 4?d trial, differences in mean daily consumption preferences of 2 varieties were separated at P‐values as small as 2 × 10^?8. The single‐elimination tournament design is an efficient means of identifying the most and least desirable varieties among a larger set of samples. One application for identifying the 2 extremes in preference within a group of varieties would be to use them as parents of a population to identify quantitative trait loci for preference.     

Comparison of two low-hazard organic solvents as individual and cosolvents for the fabrication of polysulfone membranes

Petroleum-derived solvents commonly used in membrane fabrication are often hazardous and toxic, so the investigation of safer alternatives is important. In this study, two low-hazard solvents, methyl 5-(dimethylamino)-2-methyl-5-oxopentanoate (Rhodiasolv® PolarClean) and gamma-valerolactone (GVL), were investigated as sole solvents and as cosolvents to cast polysulfone membranes via nonsolvent induced phase inversion. Normalized viscosity was introduced as an indicator of dope solution homogeneity and was used to compare the required time of mixing to achieve full dissolution of the polymer in the different solvents/solvent mixtures. All dope solutions made with low-hazard solvents were found to be more viscous than those made with traditional solvents, which meant additional mixing time was needed, and that fabricated membranes were morphologically different. With respect to operation, membranes cast from dope solutions containing equal amounts of PolarClean and GVL displayed the most similar flux curves and solute rejection to those made using the traditional solvent tested.     

Fatty acid composition of some pine seed oils

The fatty acid composition of seeds from seven species of the genusPinus (P. pinaster, P. griffithii, P. pinea, P. koraiensis, P. sylvestris, P. mughus, andP. nigra) was established. Pine seeds are rich in oil (31–68% by weight) and contain several unusual polymethylene-interrupted unsaturated fatty acids with acis-5 ethylenic bond. These are thecis-5,cis-9 18:2,cis-5,cis-9,cis-12 18:3,cis-5,cis-11 20:2, andcis-5,cis-11,cis-14 20:3 acids, with a trace ofcis-5,cis-9,cis-12,cis-15 18:4 acid. Their percentage relative to total fatty acids varies from a low of 3.1% (P. pinea) to a high of 30.3% (P. sylvestris), depending on the species. The majorcis-5 double bond-containing acid is generally thecis-5,cis-9,cis-12 18:3 acid (pinolenic acid). In all species, linoleic acid represents approximately one-half the total fatty acids, whereas the content of oleic acid varies in the range 14–36% inversely to the sum of fatty acids containing acis-5 ethylenic bond. The easily available seeds fromP. koraiensis appear to be a good source of pinolenic acid: their oil content isca. 65%, and pinolenic represents about 15% of total fatty acids. These values appear to be rather constant.Pinus pinaster, which is grown on several thousand acres in the southwest of France, is an interesting source ofcis-5,cis-11,cis-14 20:3 acid (7% in the oil, which isca. 35% of the dehulled seed weight), an acid sharing in common three double bonds with arachidonic acid. Apparently,P. sylvestris seed oil contains the highest level ofcis-5 double bond-containing acids among pine seed oils that have ever been analyzed.     

Thermal Annealing of Gold Thin Films on the Structure and Surface Morphology Using RF Magnetron Sputtering

Gold(Au)thin films were deposited on SiO2 substrate under argon(Ar)gas environment using RF(radio frequency)magnetron sputtering at room temperature for various...