Name: low-cost automated flight intercept trap for the temporal sub-sampling of flying insects attracted to artificial light at night
Uploaded: 2021-12-29T14:15:00
Description: Watch this Scientific Journal Video about Low-Cost Automated Flight Intercept Trap for the Temporal Sub-Sampling of Flying Insects Attracted to Artificial Light at Night at JoVE.com

The research was funded through the La Trobe University Net Zero Fund, sponsored by Sonepar. The research was conducted under Department of Environment, Land, Water and Planning scientific permit no. 10009741. We thank Martin Steinbauer for comments on an early draft and two anonymous reviewers.

1. Trap construction
NOTE: All of the components required to build the traps can be found in the Table of Materials. Each trap was constructed as shown in Figure 1 and Figure 2 by one person within 2 h.
<ol>
	<li>Use a jigsaw to cut the polycarbonate roofing sheets (8 mm x 610 mm x 2400 mm) into 610 mm x 230 mm sections (Figure 1, item 1 &#38; 2). Then cut a 8 mm center...

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	<li>Epsky, N. D., Morrill, W. L., Mankin, R. W., Capinera, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Traps+for+Capturing+Insects.">Traps for Capturing Insects.</a> Encyclopedia of Entomology. , (2008).</li><li>Catanach, T. A., Silvy, N. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Invertebrate+sampling+methods+for+use+in+wildlife+studies.">Invertebrate sampling methods for use in wildlife studies.</a> The Wildlife Techniques Manual. 1, 336-348 (2012).</li><li>Montgomery, G. A., Belitz, M. W., Guralnick, R. P., Tingley, M. 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J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Using+ultra-violet+traps+to+monitor+autumn+gum+moth,+Mnesampela+private+(Lepidoptera:+Geometridae),+in+south-eastern+Australia.">Using ultra-violet traps to monitor autumn gum moth, Mnesampela private (Lepidoptera: Geometridae), in south-eastern Australia.</a> Australian Forestry. 66 (4), 279-286 (2003).</li><li>Wakefield, A., Broyles, M., Stone, E. L., Harris, S., Jones, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantifying+the+attractiveness+of+broad-spectrum+street+lights+to+aerial+nocturnal+insects.">Quantifying the attractiveness of broad-spectrum street lights to aerial nocturnal insects.</a> Journal of Applied Ecology. 55, 714-722 (2017).</li><li>Williams, C. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+time+of+activity+of+certain+nocturnal+insects,+chiefly+Lepidoptera,+as+indicated+by+a+light-trap.">The time of activity of certain nocturnal insects, chiefly Lepidoptera, as indicated by a light-trap.</a> Transactions of the Entomological Society of London. 83 (4), 523-555 (1935).</li><li>Bolliger, J., Collet, M., Hohl, M., Obrist, M. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Automated+flight-interception+traps+for+interval+sampling+of+insects.">Automated flight-interception traps for interval sampling of insects.</a> PLoS ONE. 15 (7), 0229476 (2020).</li><li>Grubisic, M., van Grunsven, R. H. A., Kyba, C. C. 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J., Cemak, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+design+and+some+preliminary+results+for+a+flight+intercept+trap+to+sample+forest+canopy+arthropods.">A new design and some preliminary results for a flight intercept trap to sample forest canopy arthropods.</a> Australian Journal of Entomology. 36, 51-55 (1997).</li><li>Lamarre, G. P. A., Molto, Q., Fine, P. V. A., Baraloto, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+comparison+of+two+common+flight+interception+traps+to+survey+tropical+arthropods.">A comparison of two common flight interception traps to survey tropical arthropods.</a> ZooKeys. 216, 43-55 (2012).</li><li>Wilkening, A. J., Foltz, J. L., Atkinson, T. H., Connor, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+omnidirectional+flight+trap+for+ascending+and+descending+insects.">An omnidirectional flight trap for ascending and descending insects.</a> The Canadian Entomologist. 113, 453-455 (1981).</li><li>Frost, S. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insects+captured+in+light+traps+with+and+without+baffles.">Insects captured in light traps with and without baffles.</a> The Canadian Entomologist. 90 (9), 566-567 (1958).</li><li>Muirhead-Thompson, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Trap responses of flying insects: The influence of trap design on capture efficiency. , 287 (1991).</li><li>Carrel, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+aerial-interception+trap+for+arthropod+sampling.">A novel aerial-interception trap for arthropod sampling.</a> Florida Entomologist. 85 (4), 656-657 (2002).</li><li>Steinbauer, M. J., Haslem, A., Edwards, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Using+meteorological+and+lunar+information+to+explain+catch+variability+of+Orthoptera+and+Lepidoptera+from+250+W+Farrow+light+traps.">Using meteorological and lunar information to explain catch variability of Orthoptera and Lepidoptera from 250 W Farrow light traps.</a> Insect Conservation and Diversity. 5, 367-380 (2012).</li><li>Recher, H. F., Majer, J. D., Ganesh, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Seasonality+of+canopy+invertebrate+communities+in+eucalypt+forests+of+eastern+and+western+Australia.">Seasonality of canopy invertebrate communities in eucalypt forests of eastern and western Australia.</a> Australian Journal of Ecology. 21, 64-80 (1996).</li><li>van Klink, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Meta-analysis+reveals+declines+in+terrestrial+but+increases+in+freshwater+insect+abundances.">Meta-analysis reveals declines in terrestrial but increases in freshwater insect abundances.</a> Science. 368, 417-420 (2020).</li><li>Wagner, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insect+declines+in+the+Anthropocene.">Insect declines in the Anthropocene.</a> Annual Review of Entomology. 65, 457-480 (2020).</li><li>Cardoso, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Scientists'+warning+to+humanity+on+insect+extinctions.">Scientists' warning to humanity on insect extinctions.</a> Biological Conservation. 242, 108426 (2020).</li><li>Saunders, M. E., Janes, J. K., O'Hanlon, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Moving+on+from+the+insect+apocalypse+narrative:+Engaging+with+evidence-based+insect+conservation.">Moving on from the insect apocalypse narrative: Engaging with evidence-based insect conservation.</a> BioScience. 70 (1), 80-89 (2020).</li><li>Cardoso, P., Leather, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Predicting+a+global+insect+apocalypse.">Predicting a global insect apocalypse.</a> Insect Conservation and Diversity. 12, 263-267 (2019).</li><li>Owens, A. C. S., Cochard, P., Durrant, J., Perkin, E., Seymoure, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Light+pollution+is+a+driver+of+insect+declines.">Light pollution is a driver of insect declines.</a> Biological Conservation. 241, 108259 (2020).</li><li>Chapman, J. A., Kinghorn, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Window+traps+for+insects.">Window traps for insects.</a> The Canadian Entomologist. 87 (1), 46-47 (1955).</li><li>Canaday, C. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+insect+fauna+captured+in+six+different+trap+types+in+a+Douglas-fir+forest.">Comparison of insect fauna captured in six different trap types in a Douglas-fir forest.</a> The Canadian Entomologist. 119, 1101-1108 (2012).</li><li>Burns, M., Hancock, G., Robinson, J., Cornforth, I., Blake, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two+novel+flight-interception+trap+designs+for+low-cost+forest+insect+surveys.">Two novel flight-interception trap designs for low-cost forest insect surveys.</a> British Journal of Entomology and Natural History. 27, 155-162 (2014).</li><li>Basset, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+composite+interception+trap+for+sampling+arthropods+in+tree+canopies.">A composite interception trap for sampling arthropods in tree canopies.</a> Journal of the Australian Entomological Society. 27, 213-219 (1988).</li><li>Russo, L., Stehouwer, R., Heberling, J. 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Despite the automated flight-intercept trap described by Bolliger et al. (2020)8 being well designed and very effective at sampling at user-defined time periods, they are likely to be cost-prohibitive for many researchers. This study shows that passive trapping surveys using automated traps for sub-sampling flying insects at user-defined periods can be carried out on a modest budget. Traps were built to sample at six pre-defined time points by utilizing a commercial pet food dispenser and material...

There are many arthropod sampling techniques1,2,3, but ecologists often have difficulty applying these methods in ways that are appropriate to their research questions (see4). When choosing an appropriate method for sampling insects, ecologists must consider the targeted species, time, effort, and cost involved in different techniques. For example, a common limitation is that it can be logistically challenging to sub-sample during specific time periods over replicated sites to quantify temporal variables which influence species activity, such as changes in weather or circadian activity (but see5). Most passive-survey insect traps are set for long periods (e.g., over multiple days, weeks, or even months), lacking fine-scale temporal resolution1. For surveys targeting specific time periods across multiple replicate sites (such as nocturnal sampling only across distinct sites), a large team may be required to visit sites over multiple days at the same time points (e.g., within 30 min of sunrise and sunset) to collect specimens and reset traps6; otherwise, an automated trapping device is required5,7,8.
There is a growing field of work on the impacts of artificial light at night (ALAN) on insect activity patterns and localized population dynamics9,10; and on the interactions between ALAN and rates of insect predation4,11,12,13. However, to study the impacts of ALAN on nocturnal insect taxa, sampling needs to be confined to nighttime. Several different active light traps have been described and used for automated temporal sampling of nocturnal insects14. Some examples include simple falling disk-type separation devices, where the catch falls into a narrow tube with a disk falling every hour to separate the catch15, or turn-table separation devices that rotate collection bottles at timed intervals7,16,17. These previous automated light traps address the sampling challenges involved with temporal survey requirements but are often large and unwieldy and use outdated or unreliable technology. A new automated passive sampling device was recently developed and tested8. This device utilized a commercially available flight-interception trap paired with a lightweight custom-designed collection device consisting of a turn-table holding sampling cup that allows for collecting trap contents at user-defined intervals8. This new automated trap employs sophisticated programming that can be operated by a smartphone but is prohibitively expensive to build at around EURO 700 (AUD 1,000) per trap8.
Flight intercept traps are one of the most efficient ways to survey flying insects1,18,19 and work on the principle that flying insects fall to the ground when they collide with a vertical surface. Flight intercept traps come in a variety of designs. However, most are typically constructed with a transparent or mesh surface and a collecting container filled with water and/or a preservative. The new trap described here uses a cross vane/baffle type or multidirectional intercept trap20, given that cross baffles have been shown to increase capture rates14,21 and sample insects from all directions. The purpose of this trap is to survey nocturnal flying insects that are attracted to artificial lights. This phototaxis results in insects circling the light source22; hence a multidirectional trap is most suitable.
Described here is a low-cost automated intercept trap that requires no specialist equipment or skills to construct and operate. The trap uses a commercially available automated pet food dispenser and common items available from hardware stores. This design costs less than EURO 66 (AUD 105) per trap to construct (Table 1), making them a viable option for studies requiring temporal sub-sampling across multiple sites simultaneously.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Batteries (C cell) &#8211; 10 pack</td>
			<td>Duracell</td>
			<td>MN1400B10</td>
			<td>https://www.duracell.com.au/product/alkaline-c-batteries/</td>
		</tr>
		<tr>
			<td>Battery operated automated 6 meal pet food bowl &#8211; each</td>
			<td>OEM China</td>
			<td>XR-P006-002</td>
			<td>Automated 6-meal pet food bowls range in price dependent on supplier, for example in the UK they can be purchased for &#163;19 GBP ($36 AUD).</td>
		</tr>
		<tr>
			<td>Galvanised hex-head screws (10-16 x 16 mm) &#8211; 100 pack</td>
			<td>Bunnings Warehouse</td>
			<td>1-311-9151-CTPME</td>
			<td>Bunnings Warehouse is an Australian hardware chain with stores in Australia and New Zealand. Items purchased from Bunnings Warehouse can be found at most hardware stores. https://www.bunnings.com.au/</td>
		</tr>
		<tr>
			<td>Galvanised steel angle bracket (125 x 150 mm) &#8211; each</td>
			<td>Bunnings Warehouse</td>
			<td>AZ11</td>
			<td>https://www.bunnings.com.au/</td>
		</tr>
		<tr>
			<td>Galvanised tie wire (0.70 mm x 75 m) &#8211; per roll</td>
			<td>Bunnings Warehouse</td>
			<td>50218</td>
			<td>https://www.bunnings.com.au/</td>
		</tr>
		<tr>
			<td>Plastic basin (38 cm, 9 L round) &#8211; each</td>
			<td>Ezy Storage</td>
			<td>FBA31541</td>
			<td>https://www.ezystorage.com/product/laundry/basic-accessories/9l-round-basin/</td>
		</tr>
		<tr>
			<td>Plastic funnel (24 cm) &#8211; each</td>
			<td>Sandleford</td>
			<td>Pf24</td>
			<td>https://www.sandleford.com.au/plastic-funnel-24cm</td>
		</tr>
		<tr>
			<td>Stainless steel angle bracket (20 mm) &#8211; 16 pack</td>
			<td>Bunnings Warehouse</td>
			<td>WEB2020</td>
			<td>https://www.bunnings.com.au/</td>
		</tr>
		<tr>
			<td>Stainless steel screws &#38; nuts (M4 x 15 mm) &#8211; 18 pack</td>
			<td>Bunnings Warehouse</td>
			<td>SFA394</td>
			<td>https://www.bunnings.com.au/</td>
		</tr>
		<tr>
			<td>Stainless steel washers (3/16&#8221; &#38; M5) &#8211; 50 pack</td>
			<td>Bunnings Warehouse</td>
			<td>EBM5005</td>
			<td>https://www.bunnings.com.au/</td>
		</tr>
		<tr>
			<td>Sunlite Polycarbonate roofing sheet (8mm x 610 mm x 2.4 m) &#8211; each</td>
			<td>Suntuf (Palram Industries Ltd)</td>
			<td>SL8CL2.4</td>
			<td>https://www.palram.com/au/product/sunlite-polycarbonate-multi-wall/</td>
		</tr>
		<tr>
			<td>Treated pine paling (150 x 12 mm) &#8211; each</td>
			<td>STS Timber Wholesale P/L</td>
			<td>n/a</td>
			<td>https://www.ststimber.com.au/sts-timber-wholesale-products/fencing</td>
		</tr>
		<tr>
			<td>Wooden stakes (1200 x 17 x 17 mm) &#8211; 10 pack</td>
			<td>Lattice Makers</td>
			<td>n/a</td>
			<td>https://latticemakers.com/product/tomato-stakes-17x17mm-pack-of-10/</td>
		</tr>
	</tbody>
</table>

low-cost automated flight intercept trap for the temporal sub-sampling of flying insects attracted to artificial light at night

Sampling methods are selected depending on the targeted species or the spatial and temporal requirements of the study. However, most methods for passive sampling of flying insects have a poor temporal resolution because it is time-consuming, costly and/or logistically difficult to perform. Effective sampling of flying insects attracted to artificial light at night (ALAN) requires sampling at user-defined time points (nighttime only) across well-replicated sites resulting in major time and labor-intensive survey effort or expensive automated technologies. Described here is a low-cost automated intercept trap that requires no specialist equipment or skills to construct and operate, making it a viable option for studies that require temporal sub-sampling across multiple sites. The trap can be used to address a wide range of other ecological questions that require a greater temporal and spatial scale than is feasible with previous trap technology.

The traps were trialed in a survey of flying insects attracted to experimental lighting at four bushland reserves across Melbourne, Australia. Sites consisted of either remnant or revegetated bushland surrounded by residential housing and averaged 15 km apart (range 3-24 km) and 45 ha in size (range 30-59 ha). A total of sixteen traps were installed, four at each site, with and without experimental lights (3 lights and 1 control per site), and surveyed for 3 days and 3 nights from 30 March to 2 April 2021. Installing the...

Watch this Scientific Journal Video about Low-Cost Automated Flight Intercept Trap for the Temporal Sub-Sampling of Flying Insects Attracted to Artificial Light at Night at JoVE.com

Low-Cost Automated Flight Intercept Trap for the Temporal Sub-Sampling of Flying Insects Attracted to Artificial Light at Night

To study the impacts of artificial light at night (ALAN) on nocturnal flying insects, sampling needs to be confined to nighttime. The protocol describes a low-cost automated flight intercept trap that allows researchers to sample at user-defined periods with increased replication.

Sampling methods are selected depending on the targeted species or the spatial and temporal requirements of the study. However, most methods for passive ...

In the study of light pollution, you need to sample between dusk and dawn. Our low cost automated flight intercept traps enable the capture of flying insects at user defined intervals. Sampling multiple sites simultaneously can be difficult because you need to have researchers located at each site to start and stop your equipment simultaneously.This trap can be used in any ecological study on flying insects that requires greater temporal or spatial resolution, which cannot be provided efficiently by other sampling techniques. An example may be circadian or seasonal patterns of activity. No specialized skills are required to build this trap.You just need to be comfortable using simple workshop tools. Begin by cutting the polycarbonate roofing sheets with a jig saw. Then, cut an eight millimeter center groove halfway up the center of each pane.The two panes can now slide together to form a cross baffle. Slide the cross baffles snugly into the plastic funnel opening. Secure them to the funnel with 20 millimeter stainless steel angle brackets by drilling holes while the angle brackets are in place.Then use M4 screws and nuts with washers to secure the cross baffles to the funnel. Cut another piece of the polycarbonate sheeting from the remaining sheets. Drill holes and secure the piece at a 90 degree angle to the top of the cross baffles to form a protective roof using 20 millimeter stainless steel angle brackets, M4 screws, nuts and washers.Trim the funnel spout to a length of approximately 30 millimeters with a hacksaw to ensure that the sample trays of the automated pet feeder will rotate unobstructed at the programmed intervals. Place the automated pet dispenser with a nine liter and 30 millimeter diameter plastic basin to protect the samples from weather conditions. Drill a 20 millimeter hole into the top of the nine liter basin and place the funnel spout into the hole to position it directly above the sample tray.Using a drill with a hex head driver bit secure the plastic basin to a 500 millimeter piece of treated pine fence paling with galvanized hex head screws. To stabilize the entire trap for hoisting into the air via ropes, attach a wooden stake to the piece of treated pine fence paling with an angle bracket and tie wire. At the sampling location, remove the automated pet food dispenser from beneath the plastic basin.Then, open up the pet food dispenser and place foil dishes containing soapy water, or a preservative of choice, in each food tray. Set the clock time and then program each pet food dispenser tray to set the food tray rotation times by following the directions provided with the automated pet food dispenser. Place the food dispenser back underneath the plastic basin.Secure the basin to the timber fence piece with the galvanized hex head screws. Attach a rope to the top of the trap with a carabiner. Then, hoist the trap into position using a ladder and secure it beneath the experimental lights by the carabiner.Attach a second wooden stake to the tree with an angle bracket to stabilize the trap in high winds. After the trap sits on top of the stake, secure it with two long cable ties. To collect the insect samples, lower the traps with a rope, remove the automated pet food dispenser from beneath the plastic basin, then remove the lid of the pet food dispenser and lift out the aluminum trays.Finally, pour the contents of the trays into the pre-labeled sample vials. A comparison of relative capture efficiency of various flight intercept traps is shown here. In the present study, a total of 488 flying insects were captured over three sampling days at four sites.Two types of traps were examined in this study. Traps placed under lights and those without experimental lights. A total of 16 traps were installed, among which 12 were with experimental lights and four were without experimental lights.The number of trap days, which is equal to the number of traps multiplied by the number of days, is 36 for the traps with experimental lights and 12 for the traps without experimental lights. Therefore, to evaluate the efficiency of the traps, the total number of arthropods collected were divided by the effective surface area of each trap and the number of trap days on which they were operated. Traps perform similarly to other multidirectional baffle style traps under lights and without lights.Hence, the traps appear to be as effective as traditional flight intercept traps, but with the added benefit of sub-sampling at user defined time periods. Each collection tray holds approximately 330 milliliters which will accommodate most applications, but it would be beneficial to test the traps during swarming events.

low-cost-automated-flight-intercept-trap-for-temporal-sub-sampling

Research

JoVE Journal

Environment

Pretreatment of Lignocellulosic Biomass with Low-cost Ionic Liquids

A number of ionic liquids (ILs) with economically attractive production costs have recently received growing interest as media for the delignification of a variety of lignocellulosic feedstocks. Here we demonstrate the use of these low-cost protic ILs in the deconstruction of lignocellulosic biomass (Ionosolv pretreatment), yielding cellulose and a purified lignin. In the most generic process, the protic ionic liquid is synthesized by accurate combination of aqueous acid and amine base. The water content is adjusted subsequently. For the delignification, the biomass is placed into a vessel with IL solution at elevated temperatures to dissolve the lignin and hemicellulose, leaving a cellulose-rich pulp ready for saccharification (hydrolysis to fermentable sugars). The lignin is later precipitated from the IL by the addition of water and recovered as a solid. The removal of the added water regenerates the ionic liquid, which can be reused multiple times. This protocol is useful to investigate the significant potential of protic ILs for use in commercial biomass pretreatment/lignin fractionation for producing biofuels or renewable chemicals and materials.

The pretreatment of lignocellulosic biomass with protic low-cost ionic liquids is shown, resulting in a delignified cellulose-rich pulp and a purified lignin. The pulp gives rise to high glucose yields after enzymatic saccharification.

A number of ionic liquids (ILs) with economically attractive production costs have recently received growing interest as media for the delignification of ...

Automated, High-resolution Mobile Collection System for the Nitrogen Isotopic Analysis of NOx

Nitrogen oxides (NOx = NO + NO2) are a family of atmospheric trace gases that have great impact on the environment. NOx concentrations directly influence the oxidizing capacity of the atmosphere through interactions with ozone and hydroxyl radicals. The main sink of NOx is the formation and deposition of nitric acid, a component of acid rain and a bioavailable nutrient. NOx is emitted from a mixture of natural and anthropogenic sources, which vary in space and time. The collocation of multiple sources and the short lifetime of NOx make it challenging to quantitatively constrain the influence of different emission sources and their impacts on the environment. Nitrogen isotopes of NOx have been suggested to vary amongst different sources, representing a potentially powerful tool to understand the sources and transport of NOx. However, previous methods of collecting atmospheric NOx integrate over long (week to month) time spans and are not validated for the efficient collection of NOx in relevant, diverse field conditions. We report on a new, highly efficient field-based system that collects atmospheric NOx for isotope analysis at a time resolution between 30 min and 2 hr. This method collects gaseous NOx in solution as nitrate with 100% efficiency under a variety of conditions. Protocols are presented for collecting air in urban settings under both stationary and mobile conditions. We detail the advantages and limitations of the method and demonstrate its application in the field. Data from several deployments are shown to 1) evaluate field-based collection efficiency by comparisons with in situ NOx concentration measurements, 2) test the stability of stored solutions before processing, 3) quantify in situ reproducibility in a variety of urban settings, and 4) demonstrate the range of N isotopes of NOx detected in ambient urban air and on heavily traveled roadways.

Automated, High-resolution Mobile Collection System for the Nitrogen Isotopic Analysis of NOx

Previous work suggests that the nitrogen isotopic composition of atmospheric nitrogen oxides might distinguish the influence of different sources in the environment. We report on an automated, mobile, field-based method for the high collection efficiency of atmospheric NOx for N isotopic analysis at an hourly time resolution.

Nitrogen oxides (NOx = NO + NO2) are a family of atmospheric trace gases that have great impact on the environment. NOx concentrations directly influence ...

Methodology for Developing Life Tables for Sessile Insects in the Field Using the Whitefly, Bemisia tabaci, in Cotton As a Model System

Life tables provide a means of measuring the schedules of birth and death from populations over time. They also can be used to quantify the sources and rates of mortality in populations, which has a variety of applications in ecology, including agricultural ecosystems. Horizontal, or cohort-based, life tables provide for the most direct and accurate method of quantifying vital population rates because they follow a group of individuals in a population from birth to death. Here, protocols are presented for conducting and analyzing cohort-based life tables in the field that takes advantage of the sessile nature of the immature life stages of a global insect pest, Bemisia tabaci. Individual insects are located on the underside of cotton leaves and are marked by drawing a small circle around the insect with a non-toxic pen. This insect can then be observed repeatedly over time with the aid of hand lenses to measure development from one stage to the next and to identify stage-specific causes of death associated with natural and introduced mortality forces. Analyses explain how to correctly measure multiple mortality forces that act contemporaneously within each stage and how to use such data to provide meaningful population dynamic metrics. The method does not directly account for adult survival and reproduction, which limits inference to dynamics of immature stages. An example is presented that focused on measuring the impact of bottom-up (plant quality) and top-down (natural enemies) effects on the mortality dynamics of B. tabaci in the cotton system.

Methodology for Developing Life Tables for Sessile Insects in the Field Using the Whitefly, Bemisia tabaci, in Cotton As a Model System

Life tables allow quantification of the sources and rates of mortality in insect populations and contribute to understanding, predicting and manipulating population dynamics in agroecosystems. Methods for conducting and analyzing cohort-based life tables in the field for an insect with sessile immature life stages are presented.

Life tables provide a means of measuring the schedules of birth and death from populations over time. They also can be used to quantify the sources and ...

Adherence of Bacteria to Plant Surfaces Measured in the Laboratory

This manuscript describes a method to measure bacterial binding to axenic plant surfaces in the light microscope and through the use of viable cell counts. Plant materials used include roots, sprouts, leaves, and cut fruits. The methods described are inexpensive, easy, and suitable for small sample sizes. Binding is measured in the laboratory and a variety of incubation media and conditions can be used. The effect of inhibitors can be determined. Situations that promote and inhibit binding can also be assessed. In some cases it is possible to distinguish whether various conditions alter binding primarily due to their effects on the plant or on the bacteria.

An easy method for measuring and characterizing bacterial adhesion to plants, particularly roots and sprouts, is described in this article.

This manuscript describes a method to measure bacterial binding to axenic plant surfaces in the light microscope and through the use of viable cell ...

A Strain Gauge Monitor (SGM) for Continuous Valve Gape Measurements in Bivalve Molluscs in Response to Laboratory Induced Diel-cycling Hypoxia and pH

An inexpensive, laboratory-based, strain gauge valve gape monitor (SGM) was developed to monitor the valve gape behavior of bivalve molluscs in response to diel-cycling hypoxia. A Wheatstone bridge was connected to strain gauges that were attached to the shells of oysters (Crassostrea virginica). The recorded signals allowed for the opening and closing of the bivalves to be recorded continuously over two-day periods of experimentally-induced diel-cycling hypoxia and diel-cycling changes in pH. Here, we describe a protocol for developing an inexpensive strain gauge monitor and describe, in an example laboratory experiment, how we used it to measure the valve gape behavior of Eastern oysters (C. virginica), in response to diel-cycling hypoxia and cyclical changes in pH. Valve gape was measured on oysters subjected to cyclical severe hypoxic (0.6 mg/L) dissolved oxygen conditions with and without cyclical changes in pH, cyclical mild hypoxic (1.7 mg/L) conditions and normoxic (7.3 mg/L) conditions. We demonstrate that when oysters encounter repeated diel cycles, they rapidly close their shells in response to severe hypoxia and close with a time lag to mild hypoxia. When normoxia is restored, they rapidly open again. Oysters did not respond to cyclical pH conditions superimposed on diel cycling severe hypoxia. At reduced oxygen conditions, more than one third of the oysters closed simultaneously. We demonstrate that oysters respond to diel-cycling hypoxia, which must be considered when assessing the behavior of bivalves to dissolved oxygen. The valve SGM can be used to assess responses of bivalve molluscs to changes in dissolved oxygen or contaminants. Sealing techniques to better seal the valve gape strain gauges from sea water need further improvement to increase the longevity of the sensors.

A Strain Gauge Monitor SGM for Continuous Valve Gape Measurements in Bivalve Molluscs in Response to Laboratory Induced Diel-cycling Hypoxia and pH

Understanding the behavioral responses of bivalve suspension-feeders to environmental variables, such as dissolved oxygen, can explain some ecosystem processes. We have developed an inexpensive, laboratory-based, strain gauge monitor (SGM) to measure valve gape responses of oysters, Crassostrea virginica, to diel-cycling hypoxia and cyclical pH.

An inexpensive, laboratory-based, strain gauge valve gape monitor (SGM) was developed to monitor the valve gape behavior of bivalve molluscs in response ...

Measuring the Flight Ability of the Ambrosia Beetle, Platypus Quercivorus (Murayama), Using a Low-Cost, Small, and Easily Constructed Flight Mill

The ambrosia beetle, Platypus quercivorus (Murayama), is the vector of a fungal pathogen that causes mass mortality of Fagaceae trees (Japanese oak wilt). Therefore, knowing the dispersal capacity may help inform trapping/tree removal efforts to prevent this disease more effectively. In this study, we measured the flight velocity and duration and estimated the flight distance of the beetle using a newly developed flight mill. The flight mill is low cost, small, and constructed using commonly available items. Both the flight mill arm and its vertical axis comprise a thin needle. A beetle specimen is glued to one tip of the arm using instant glue. The other tip is thick due to being covered with plastic, thus it facilitates the detection of rotations of the arm. The revolution of the arm is detected by a photo sensor mounted on an infrared LED, and is indicated by a change in the output voltage when the arm passed above the LED. The photo sensor is connected to a personal computer and the output voltage data are stored at a sampling rate of 1 kHz. By conducting experiments using this flight mill, we found that P. quercivorus can fly at least 27 km. Because our flight mill comprises cheap and small ordinary items, many flight mills can be prepared and used simultaneously in a small laboratory space. This enables experimenters to obtain a sufficient amount of data within a short period.

Measuring the Flight Ability of the Ambrosia Beetle, Platypus Quercivorus Murayama, Using a Low-Cost, Small, and Easily Constructed Flight Mill

We developed a low cost and small flight mill, constructed with commonly available items and easily used in experimentation. Using this apparatus, we measured the flight ability of an ambrosia beetle, Platypus quercivorus.

The ambrosia beetle, Platypus quercivorus (Murayama), is the vector of a fungal pathogen that causes mass mortality of Fagaceae trees (Japanese oak wilt). ...

Harvesting Venom Toxins from Assassin Bugs and Other Heteropteran Insects

Heteropteran insects such as assassin bugs (Reduviidae) and giant water bugs (Belostomatidae) descended from a common predaceous and venomous ancestor, and the majority of extant heteropterans retain this trophic strategy. Some heteropterans have transitioned to feeding on vertebrate blood (such as the kissing bugs, Triatominae; and bed bugs, Cimicidae) while others have reverted to feeding on plants (most Pentatomomorpha). However, with the exception of saliva used by kissing bugs to facilitate blood-feeding, little is known about heteropteran venoms compared to the venoms of spiders, scorpions and snakes.
One obstacle to the characterization of heteropteran venom toxins is the structure and function of the venom/labial glands, which are both morphologically complex and perform multiple biological roles (defense, prey capture, and extra-oral digestion). In this article, we describe three methods we have successfully used to collect heteropteran venoms. First, we present electrostimulation as a convenient way to collect venom that is often lethal when injected into prey animals, and which obviates contamination by glandular tissue. Second, we show that gentle harassment of animals is sufficient to produce venom extrusion from the proboscis and/or venom spitting in some groups of heteropterans. Third, we describe methods to harvest venom toxins by dissection of anaesthetized animals to obtain the venom glands. This method is complementary to other methods, as it may allow harvesting of toxins from taxa in which electrostimulation and harassment are ineffective. These protocols will enable researchers to harvest toxins from heteropteran insects for structure-function characterization and possible applications in medicine and agriculture.

Although many insects in the suborder Heteroptera (Insecta: Hemiptera) are venomous, their venom composition and the functions of their venom toxins are mostly unknown. This protocol describes methods to harvest heteropteran venoms for further characterization, using electrostimulation, harassment, and gland dissection.

Heteropteran insects such as assassin bugs (Reduviidae) and giant water bugs (Belostomatidae) descended from a common predaceous and venomous ancestor, ...

A Flexible Low Cost Hydroponic System for Assessing Plant Responses to Small Molecules in Sterile Conditions

A wide range of studies in plant biology are performed using hydroponic cultures. In this work, an in vitro hydroponic growth system designed for assessing plant responses to chemicals and other substances of interest is presented. This system is highly efficient in obtaining homogeneous and healthy seedlings of the C3 and C4 model species Arabidopsis thaliana and Setaria viridis, respectively. The sterile cultivation avoids algae and microorganism contamination, which are known limiting factors for plant normal growth and development in hydroponics. In addition, this system is scalable, enabling the harvest of plant material on a large scale with minor mechanical damage, as well as the harvest of individual parts of a plant if desired. A detailed protocol demonstrating that this system has an easy and low-cost assembly, as it uses pipette racks as the main platform for growing plants, is provided. The feasibility of this system was validated using Arabidopsis seedlings to assess the effect of the drug AZD-8055, a chemical inhibitor of the target of rapamycin (TOR) kinase. TOR inhibition was efficiently detected as early as 30 min after an AZD-8055 treatment in roots and shoots. Furthermore, AZD-8055-treated plants displayed the expected starch-excess phenotype. We proposed this hydroponic system as an ideal method for plant researchers aiming to monitor the action of plant inducers or inhibitors, as well as to assess metabolic fluxes using isotope-labeling compounds which, in general, requires the use of expensive reagents.

A simple, versatile, and low-cost in vitro hydroponic system was successfully optimized, enabling large-scale experiments under sterile conditions. This system facilitates the application of chemicals in a solution and their efficient absorption by roots for molecular, biochemical, and physiological studies.

A wide range of studies in plant biology are performed using hydroponic cultures. In this work, an in vitro hydroponic growth system designed for ...

Construction of a Compact Low-Cost Radiation Shield for Air-Temperature Sensors in Ecological Field Studies

Low cost temperature sensors are increasingly used by ecologists to assess climatic variation and change on ecologically relevant scales. Although cost-effective, if not deployed with proper solar radiation shielding, the observations recorded from these sensors will be biased and inaccurate. Manufactured radiation shields are effective at minimizing this bias, but are expensive compared to the cost of these sensors. Here, we provide a detailed methodology for constructing a compact version of a previously described custom fabricated radiation shield, which is more accurate than other published shielding methods that attempt to minimize shield size or construction costs. The method requires very little material: corrugated plastic sheets, aluminum foil duct tape, and cable ties. One 15 cm and two 10 cm squares of corrugated plastic are used for each shield. After cutting, scoring, taping and stapling of the sheets, the 10 cm squares form the bottom two layers of the solar radiation shield, while the 15 cm square forms the top layer. The three sheets are held together with cable ties. This compact solar radiation shield can be suspended, or placed against any flat surface. Care must be taken to ensure that the shield is completely parallel to the ground to prevent direct solar radiation from reaching the sensor, possibly causing increased warm biases in sun-exposed sites in the morning and afternoon relative to the original, larger design. Even so, differences in recorded temperatures between the smaller, compact shield design and the original design were small (mean daytime bias = 0.06 &#176;C). Construction costs are less than half of the original shield design, and the new design results in a less conspicuous instrument that may be advantageous in many field ecology settings.

With the advent of small, low-cost environmental sensors, it is now possible to deploy high-density networks of sensors to measure hyper localized temperature variation. Here, we provide a detailed methodology for constructing a compact version of a previously described custom-fabricated radiation shield for use with inexpensive thermochrons.

Low cost temperature sensors are increasingly used by ecologists to assess climatic variation and change on ecologically relevant scales. Although ...

Fabrication of Anisotropic Polymeric Artificial Antigen Presenting Cells for CD8+ T Cell Activation

Artificial antigen presenting cells (aAPC) are a promising platform for immune modulation due to their potent ability to stimulate T cells. Acellular substrates offer key advantages over cell-based aAPC, including precise control of signal presentation parameters and physical properties of the aAPC surface to modulate its interactions with T cells. aAPC constructed from anisotropic particles, particularly ellipsoidal particles, have been shown to be more effective than their spherical counterparts at stimulating T cells due to increased binding and larger surface area available for T cell contact, as well as reduced nonspecific uptake and enhanced pharmacokinetic properties. Despite increased interest in anisotropic particles, even widely accepted methods of generating anisotropic particles such as thin-film stretching can be challenging to implement and use reproducibly.
To this end, we describe a protocol for the rapid, standardized fabrication of biodegradable anisotropic particle-based aAPC with tunable size, shape, and signal presentation for T cell expansion ex vivo or in vivo, along with methods to characterize their size, morphology, and surface protein content, and to assess their functionality. This approach to fabricating anisotropic aAPC is scalable and reproducible, making it ideal for generating aAPC for &#34;off-the-shelf&#34; immunotherapies.

Here, we present a protocol to quickly and reproducibly generate biologically inspired, biodegradable articifical antigen presenting cells (aAPC) with tunable size, shape, and surface protein presentation for T cell expansion ex vivo or in vivo.

Artificial antigen presenting cells (aAPC) are a promising platform for immune modulation due to their potent ability to stimulate T cells. Acellular ...