An engineering approach yields a highly potent combination of four antinematode drugs at lower dosages than individual drugs.
We developed an open microfluidic system to dispense and manipulate discrete droplets on planar plastic sheets. Here, a superhydrophobic material is spray-coated on commercially-available plastic sheets followed by the printing of hydrophilic symbols using an inkjet printer. The patterned plastic sheets are taped to a two-axis tilting platform, powered by stepper motors, that provides mechanical agitation for droplet transport. We demonstrate the following droplet operations: transport of droplets of different sizes, parallel transport of multiple droplets, merging and mixing of multiple droplets, dispensing of smaller droplets from a large droplet or a fluid reservoir, and one-directional transport of droplets. As a proof-of-concept, a colorimetric assay is implemented to measure the glucose concentration in sheep serum. Compared to silicon-based digital microfluidic devices, we believe that the presented system is appealing for various biological experiments because of the ease of altering design layouts of hydrophilic symbols, relatively faster turnaround time in printing plastic sheets, larger area to accommodate more tests, and lower operational costs by using off-the-shelf products.
Plant-parasitic nematodes cause substantial damage to agricultural crops worldwide. Long-term management of these pests requires novel strategies to reduce infection of host plants. Disruption of nematode chemotaxis to root systems has been proposed as a potential management approach, and novel assays are needed to test the chemotactic behavior of nematodes against a wide range of synthetic chemicals and root exudates. Two microfluidic chips were developed that measure the attraction or repulsion of nematodes to chemicals ("chemical chip") and young plant roots ("root chip"). The chip designs allowed for chemical concentration gradients to be maintained up to 24 h, the nematodes to remain physically separate from the chemical reservoirs, and for images of nematode populations to be captured using either a microscope or a flatbed scanner. In the experiments using the chemical chips, seven ionic solutions were tested on second-stage juveniles (J2s) of Meloidogyne incognita and Heterodera glycines. Results were consistent with previous reports of repellency of M. incognita to a majority of the ionic solutions, including NHNO, KNO, KCl, MgCl, and CaCl. H. glycines was found to be attracted to both NHNO and KNO, which has not been reported previously. A software program was written to aid in monitoring the location of nematodes at regular time intervals using the root chip. In experiments with the root chip, H. glycines J2s were attracted to roots of 3-day-old, susceptible (cultivar Williams 82) soybean seedlings, and attraction of H. glycines to susceptible soybean was similar across the length of the root. Attraction to resistant (cultivar Jack) soybean seedlings relative to the water only control was inconsistent across runs, and H. glycines J2s were not preferentially attracted to the roots of resistant or susceptible cultivars when both were placed on opposite sides of the same root chip. The chips developed allow for direct tests of plant-parasitic nematode chemotaxis to chemicals and roots with minimal human intervention.
In nature, several organisms possess a magnetic compass to navigate or migrate them to desired locations. It is thought that these organisms may use biogenic magnetic matter or light-sensitive photoreceptors to sense and orient themselves in magnetic fields. To unravel the underlying principles of magnetosensitivity and magnetoreception, previous experiments have been conducted on bacteria, vertebrates, crustaceans, and insects. In this study, the model organism, C. elegans, is used to test their response and sensitivity to static magnetic fields in the range of 5 milli Tesla to 120 milli Tesla. Single wild-type C. elegans are put in microfluidic channels and exposed to permanent magnets for five cycles of thirty-second time intervals. The worm movement is recorded and analyzed with custom software to calculate the average velocity and the percentage of turning and curling. Contrary to some published studies, our results did not show a significant difference compared to control experiments. This suggests that C. elegans may not sense static magnetic fields in the range of field strengths that we tested.
The objective of this study was to determine the effects of ILeVO (fluopyram) and VOTiVO (Bacillus firmus I-1582) seed treatments on Heterodera glycines second-stage juvenile (J2) root penetration and behavior. In a growth chamber experiment, roots of soybeans grown from treated or untreated seeds were inoculated with H. glycines J2s at soil depths of 2.5, 5, or 7.5 cm. ILeVO significantly reduced H. glycines root penetration compared with the untreated control, but only when J2s were inoculated at a soil depth of 2.5 cm, which was near the seed. Changes in nematode behavior were assessed by collecting 60-s videos of J2s after 2 h of exposure to exudates from treated seeds or radicles from treated seeds or from soil leachates in which treated seeds were planted. X- and y-coordinates of each of the 13 reference points were recorded every hour for 24 h. A custom program analyzed and transformed the coordinates into nematode motion parameters (speed and total change in curvature). ILeVO, but not VOTiVO, seed exudates significantly reduced J2 speed relative to the untreated control. Soil leachates from ILeVO or VOTiVO treatments had no consistent effect on H. glycines speed or total change in curvature compared with the untreated control. In another experiment, treated or untreated seeds were incubated in wells of 6-well tissue culture plates containing 11.5% Pluronic gel. Seeds were removed after 2 h, and approximately 50 J2s then were pipetted into each well. The plates were scanned every 60 min for 24 h, and the number of J2s in each well that moved a minimum distance of ≥300 µm was determined using another custom software program. ILeVO, but not VOTiVO, significantly reduced the movement of J2 populations relative to control wells in which no seeds were added. And wells that had seeds, treated or not, yielded significantly less J2 movement compared with the no-seed control. The results of these experiments indicate that ILeVO reduces activity on H. glycines J2s but may not affect nematodes beyond a limited area surrounding the treated seed.
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