Several technicians and graduate students at Michigan State University have helped to develop, refine and evaluate the DD trap design over the years, including Andrea Anulewicz, Robert McDonald and Nathan Siegert. We thank James Wieferich and Jeremy Lowell (MSU) for their assistance in developing the instructions for DD installation. James Wieferich and Molly Robinett (MSU) reviewed an earlier draft of this manuscript and we appreciate their suggestions. Joseph Francese and Damon Crook (USDA APHIS) generously shared their observations on EAB response to color and host volatiles. Funding for DD trap development and evaluation was provided by grants from the USDA Forest Service, Northeastern Area, Forest Health Protection.

1. Prepare the Green and Purple Panels
<ol>
	<li>Acquire green and purple corrugated plastic panels (120 cm &#215; 60 cm) for EAB trapping from a commercial distributor of pest management supplies. Use a box-cutter or utility knife to score two fold lines in each panel by partially cutting through the plastic along the vertical corrugations, 40 cm from the two short edges of the panel. This allows the panel to be folded into a 3-sided prism (each face will be 40 x 60 cm). Panels are usually scored by the distributor to facilitate folding, so this step may not be necessary.</li>
	<li>Drill two pairs of holes into each panel using a &#189; inch (1.3 cm) bit as indicated in Figure 1. Drill holes 4 inches (10 cm) from the top of the panel and 4 inches on either side of the left fold. Drill the second pair of holes 19 inches (48 cm) from the top of the panel and directly below the top holes (i.e., 4 inches on either side of the left fold) .</li>
</ol>
2. Prepare the PVC Pipe
<ol>
	<li>Obtain one section of schedule 40 polyvinyl chloride (PVC) pipe, 3 m long and 10 cm in diameter, from a commercial hardware store or other supplier for each trap. Drill four holes into the PVC pipe, as shown in Figure 2. Using a &#189; inch (1.3 cm) drill bit, drill a pair of holes 4 inches (10 cm) from the top of the PVC pipe and 3&#189; inches (8.9 cm) apart. Align the holes horizontally.</li>
	<li>Drill a second pair of holes, 3&#189; inches apart, and 52 inches (1.3 m) below the top of the pipe, directly below the first set of holes .</li>
</ol>
3. Install a T-post
<ol>
	<li>Use a post pounder to install a t-post in the desired location. Drive the post deep enough so that the flange is just below the soil surface, but there is no need to go any deeper. The t-post will provide plenty of support for the DD trap, even in high winds. Guy wires or additional supports are not necessary. Although the t-post does not need to be set deeply into the soil, it is a good idea to check for underground utilities or buried pipes before driving the post into the ground.</li>
</ol>
4. Assemble the DD Trap
<ol>
	<li>Fold a purple panel into a prism. If panels are pre-glued, be sure the sticky side is on the outside. Feed the two small flaps through the slits and use two 8 inch (20 cm) cable ties to fasten the panel ends together. Repeat with a green panel.</li>
	<li>Support the PVC pipe horizontally above the ground. Slide the purple prism onto the PVC pipe and down to the lower set of holes in the PVC pipe.</li>
	<li>Feed a large, 2 foot (60 cm) long cable tie horizontally through the top left hole in the prism, then through the pair of holes in the PVC pipe, and finally through the adjacent hole on the right side of the prism. Be sure the prism is snug against the solid portion of PVC pipe that lies between the two holes. Close and tighten the cable tie, making sure the cable tie is not twisted.</li>
	<li>Feed a second large cable tie through the bottom hole in the prism, around the pipe and through the corresponding hole on the adjacent side of the prism. Close the cable tie and tighten, again taking care to ensure the cable tie does not twist before it is tightened.</li>
	<li>Slide the green prism onto the PVC pipe and attach it to the upper set of holes in the PVC pipe with two 60-cm cable ties, following the same steps as before. There will be a 21 inch (54 cm) gap between the upper and lower prisms.</li>
</ol>
5. Apply Insect Glue and Set Up the Trap
<ol>
	<li>Apply clear insect trapping glue to panels that are not pre-glued. Even pre-glued panels often require additional glue to capture EAB beetles. To apply insect trapping glue, set the base of the PVC pipe on the ground and prop the inner face of the upper prism on the t-post.</li>
	<li>Using latex (or similar) gloves, scoop up two to three handfuls of glue and smear it across the surface of a panel, making sure the surface of the panel is thoroughly coated. Rotate the PVC pipe to access all three surfaces on both prisms. Glue does not need to be applied to the PVC pipe.</li>
	<li>Pick up the PVC pipe, hold it vertically and slide the lower end over the t post. Avoid touching the sticky panels.</li>
</ol>
6. Bait the DD Trap
<ol>
	<li>As pre-glued prisms usually arrive with a small hole (4 mm) near the bottom edge of one panel of the prism, use a 4 inch (10 cm) cable tie to attach one pouch of cis-3-hexenol to the lower edge of the top (green) prism. If a hole is not present, use a handheld hole punch or a pocketknife to cut a small hole or slit into the plastic, then use the small cable tie to attach the lure. 
	NOTE: The cis-3-hexenol compound in the lures is slowly released over about a 10 week period. If traps need to remain effective for more than 10 weeks, replace the lures as needed.</li>
</ol>
7. Checking Traps for EAB Beetles
<ol>
	<li>Check traps at 2-4 week intervals if possible, to avoid losing captured beetles, which occasionally drop off the traps, especially if heavy rain occurs. Examine all three sides of the lower prism for EAB beetles. Then, using disposable latex gloves, lift the PVC pipe straight up and off the t-post and prop the inner face of the upper prism on the t-post, exposing one panel of both prisms.</li>
	<li>Use forceps to remove any suspect EAB beetles and place them in a re-sealable plastic specimen bag.</li>
	<li>Rotate the PVC pipe to examine all three panels on both prisms for suspect EAB beetles, then slide the pipe back over the t-post. 
	NOTE: Flies, click beetles and various other insects will inevitably be captured on the traps. It takes a keen eye to spot an EAB beetle. Carrying a photo reference card can be helpful when checking traps for the first time.</li>
	<li>To confirm species identification, return suspect beetles to the lab and soak them in a non-toxic histological clearing agent for a day or two to remove the insect trapping glue. Beetles can then be examined under a dissecting microscope.</li>
</ol>

The authors declare that they have no competing financial interests.

Both the design and placement of DD traps exploit the attraction of adult EAB beetles to specific shades of color and to light. The green prism on the top of the PVC pipe is most attractive to male beetles, who spend their life span feeding on ash leaves, as well as mating32,36,38,39. The lower purple prism ensures traps are also attractive to female beetles32. Like male...

The emerald ash borer (EAB) (Agrilus planipennis Fairmaire) (Coleoptera: Buprestidae) has killed hundreds of millions of ash (Fraxinus spp.) trees since it was first identified in 2002 in the greater Detroit, Michigan metropolitan area and in nearby Windsor, Ontario, Canada. Inadvertent spread of infested ash trees, logs and firewood, along with natural dispersal of beetles, have resulted in EAB establishment in at least 27 states, and two Canadian provinces to date1. Recent reports indicate the EAB has also invaded Moscow, Russia, where it is killing landscape ash trees2,3, generating additional concern about its potential spread into Europe. Interspecific variation in EAB host preference and resistance among North American ash species has been documented4,5,6,7,8,9, but virtually all ash species in North America are likely to be suitable hosts. Catastrophic levels of ash mortality have been recorded in areas of Michigan and Ohio10,11,12, with associated ecological and economic impacts13,14,15,16.
Effective methods to detect new EAB infestations and monitor low density populations are critical aspects of managing this invasive pest in urban, residential and forested settings. Early detection provides time to develop a strategy, secure funding and implement activities to reduce impacts of the EAB. For example, municipal officials and homeowners can begin treating valuable ash in landscapes with systemic insecticides before damage caused by increasing larval densities limits efficacy of these products17,18. Similarly, identification of a new infestation and reliable information about local EAB distribution gives foresters and property owners the opportunity to implement timber sales, species conversion or other activities to reduce economic costs or ecological effects of ash mortality.
Early detection, delimitation, and effective monitoring of low-density EAB populations, however, remain difficult. Visual surveys to identify newly infested ash trees are not reliable because ash rarely exhibit external signs or symptoms of EAB infestation until larval densities build to moderate or even high levels4,19. The most effective means of detecting very low densities of EAB involve using girdled ash detection trees19,20,21,22. Ash trees are girdled in spring or early summer by removing a band of outer bark and phloem around the circumference of the trunk, which stresses trees, increasing their attraction to adult EAB beetles. Girdled trees can be debarked in autumn or winter to identify EAB larval presence and density. Although girdled ash trees have been used operationally for EAB detection19,23,24,25, they are problematic. Debarking girdled trees can be labor intensive and locating suitable trees for girdling can be difficult, especially in urban or residential areas or when surveys must be conducted for multiple years19.
Artificial traps baited with EAB attractants eliminate many concerns associated with using girdled ash detection trees. In contrast to other important forest pests such as gypsy moth (Lymantria dispar L) and some Scolytinae bark beetles that produce long distance sex or aggregation pheromones, to date, no effective long-distance pheromones have been found for EAB. A short range sex pheromone, cis-lactone, may facilitate mating26,27, but in field trials, cis-lactone lures have not consistently increased EAB attraction to artificial traps28. Adult beetles rely on volatile compounds emitted by ash leaves, bark and wood to identify their host trees29,30,31 and encounter potential mates. Several volatile compounds have been evaluated for use in lures to attract adult EABs to artificial traps27,32. Currently, traps used operationally for EAB detection surveys in the U.S. are baited with lures containing cis-3-hexenol, a common green leaf volatile produced by ash foliage30,33. In previous years, EAB traps used for U.S. surveys have also been baited with Manuka oil, which is extracted from the New Zealand tea tree (Leptospermum scoparium Forst and Forst ) or Phoebe oil, an extract of the Brazilian walnut tree (Phoebe porosa Mez.); both contain several sesquiterpenes that are also present in ash bark29. Problems with inconsistent supplies of these natural oils, however, have restricted their use.
In addition to host-produced volatiles, adult EAB beetles respond to visual stimuli, including color and light20,32,34,35. Early studies showed EAB adults, which are relatively agile fliers, were rarely captured by black funnel traps baited with various ash volatiles (DGM and TMP, unpublished data). Other trap designs, such as cross-vane traps, were evaluated but the aversion of EAB beetles to dark spaces and shadows limited the effectiveness of these traps.
Development of a three-sided prism, which can be coated with clear insect trapping glue35 to capture beetles, was a substantial improvement in trap design. Attraction of adult EABs to color has also been extensively evaluated in field trials and in laboratory studies with a retinograph34. Results show EAB beetles are consistently attracted to specific shades of green and purple32,36. Prism traps fabricated from colored corrugated plastic are now widely used in EAB survey activities in the U.S. and Canada.
Because EAB adults are strongly attracted to light, beetles are much more likely to colonize open-grown trees than shaded trees20,21. Guidelines for EAB detection surveys in the U.S. required individual prism traps to be suspended from a mid-canopy branch in an ash tree growing along a road or the edge of a wooded area37. In theory, this should ensure that at least one panel of the prism is exposed to sunlight. Operationally, however, prism traps may be partially shadowed by overhead branches or by adjacent or nearby trees. Sticky panel surfaces are frequently blown into foliage, resulting in leaves adhering to and obscuring at least a portion of one or more panels.
Double-decker (DD) traps were developed to integrate multiple visual and olfactory cues to enhance attraction of EAB beetles. Each DD trap is comprised of one green and one purple corrugated plastic prism attached to a 3 m tall schedule 40 polyvinyl chloride (PVC) pipe (10 cm diameter), which is supported by sliding the PVC pipe over a t-post. Using both green and purple prisms is designed to attract both sexes of EAB beetles32,36,38,39. Additionally, rather than being suspended from a branch in the canopy of an ash tree, the DD traps are placed in full sun, 5-10 m away from ash trees along the edge of a wooded area or in the midst of scattered, open-grown ash trees.

<table><tbody><tr><td>Light green corrugated plastic panel: 120 cm x 60 cm</td><td>Great Lakes IPM; www.greatlakesipm.com</td><td>IPM-EAB GR</td><td>All three surfaces of each prism need to be covered with clear insect trapping glue, even if the panels are pre-glued. Pre-glued panels are often not sticky enough to consistently capture or retain EAB beetles.&amp;nbsp; Other clear insect trapping glue products are available but are considerably more difficult to apply.&amp;nbsp;&amp;nbsp;&amp;nbsp;</td></tr><tr><td>Light purple corrugated plastic panel: 120 cm x 60 cm</td><td>Great Lakes IPM; www.greatlakesipm.com</td><td>IPM-EAB&amp;nbsp; LP</td><td></td></tr><tr><td>Large cable tie (4): 60 cm with a 79 kg capacity</td><td>Cabletiesandmore.com; http://www.cabletiesandmore.com/cableties.php</td><td>CT-24-NU-100PK</td><td></td></tr><tr><td>Medium cable ties (4): 20 cm with a 22.7 kg capacity</td><td>Cabletiesandmore.com; http://www.cabletiesandmore.com/cableties.php</td><td>CT261</td><td></td></tr><tr><td>Small cable tie: 10 cm with a 8.2 kg capacity</td><td>Cabletiesandmore.com; http://www.cabletiesandmore.com/cableties.php</td><td>CT204</td><td></td></tr><tr><td>cis-3-hexanol pouch</td><td>Synergy Semiochemicals; http://www.semiochemical.com/html/buprestids.html)</td><td>3136</td><td>Lures used to bait DD traps consist of pouches containing cis-3-hexenol, a non-toxic compound present in ash leaves.&amp;nbsp; One pouch is attached to the lower edge of the top prism using a small cable tie.&amp;nbsp; Each pouch of cis-3-hexenol has a release rate of approximately 50 mg/day. Note that cis-3-hexenol is sometimes written as Z-3-hexenol.&amp;nbsp;</td></tr><tr><td>Aphinity Hexenol</td><td>Sylvar Technologies</td><td></td><td></td></tr><tr><td>Lure GLV4 emerald ash borer</td><td>Chemtica, Heredia, Costa Rica</td><td></td><td></td></tr><tr><td>cis-3-hexanol pouch</td><td>WestGreen Global Technologies; http://www.westgreenglobaltechnologies.com/</td><td></td><td></td></tr><tr><td>Clear insect trapping glue&amp;nbsp;</td><td>Hummert International; http://www.hummert.com/product-details/8196/pestick</td><td>01-3522-1&amp;nbsp;&amp;nbsp;</td><td></td></tr><tr><td>Histoclear II histological clearing agent</td><td>National Diagnostics; www.nationaldiagnostics.com</td><td>HS-202</td><td>Histoclear II will be needed to remove the sticky insect glue from suspect beetles.&amp;nbsp; Other histological clearing agents are available but may not remove the glue and some products dissolve plastic, an important consideration if plastic containers are used for soaking the beetles.&amp;nbsp;</td></tr><tr><td>Histoclear II histological clearing agent</td><td>Great Lakes IPM; www.greatlakesipm.com</td><td>10011</td><td>Histoclear II will be needed to remove the sticky insect glue from suspect beetles.&amp;nbsp; Other histological clearing agents are available but may not remove the glue and some products dissolve plastic, an important consideration if plastic containers are used for soaking the beetles.&amp;nbsp;</td></tr><tr><td>t-post: 1.5 m</td><td>multiple sources</td><td></td><td>A t-post (5 feet tall) (1.5 m) is used to support the PVC pipe.&amp;nbsp;&amp;nbsp;</td></tr><tr><td>post pounder</td><td>multiple sources</td><td></td><td>Use a post pounder to set t-posts into the ground. No additional support is necessary.</td></tr><tr><td>HDPE (high density polyethylene) PVC pipe : 3 m x 10 cm diameter</td><td>multiple sources</td><td></td><td></td></tr><tr><td>Forceps (rigid)</td><td>multiple sources</td><td></td><td>Forceps (tweezers) will be needed to remove suspect beetles from the traps. Rigid forceps work better than flexible forceps.&amp;nbsp;</td></tr><tr><td>Latex gloves</td><td>multiple sources</td><td></td><td>Latex gloves are needed for applying the insect trapping glue to the prisms and for checking the traps to collect EAB beetles.&amp;nbsp;&amp;nbsp;&amp;nbsp;</td></tr><tr><td>Baby oil or baby wipes&amp;nbsp;</td><td>multiple sources</td><td></td><td>Baby oil or baby wipes are helpful for removing the trapping glue from hands and equipment.&amp;nbsp;</td></tr><tr><td>Re-sealable plastic specimen bags: 5 cm x 8 cm&amp;nbsp;</td><td>multiple sources</td><td></td><td>Small re-sealable plastic specimen bags are useful for collecting beetles from traps.&amp;nbsp; Each bag should be labelled, either with pre-made, adhesive labels or with soft felt pens.&amp;nbsp;&amp;nbsp;&amp;nbsp;</td></tr><tr><td>Guides to help with distinguishing EAB from beetles native to North America are available on the national EAB website at www.emeraldashborer.info. &amp;nbsp;</td><td></td><td></td><td></td></tr></tbody></table>

building double-decker traps for early detection of emerald ash borer

Emerald ash borer (EAB) (Agrilus planipennis Fairmaire), the most destructive forest insect to have invaded North America, has killed hundreds of millions of forest and landscape ash (Fraxinus spp.) trees. Several artificial trap designs to attract and capture EAB beetles have been developed to detect, delineate, and monitor infestations. Double-decker (DD) traps consist of two corrugated plastic prisms, one green and one purple, attached to a 3 m tall polyvinyl chloride (PVC) pipe supported by a t-post. The green prism at the top of the PVC pipe is baited with cis-3-hexenol, a compound produced by ash foliage. Surfaces of both prisms are coated with sticky insect glue to capture adult EAB beetles. Double-decker traps should be placed near ash trees but in open areas, exposed to sun. Double-decker trap construction and placement are presented here, along with a summary of field experiments demonstrating the efficacy of DD traps in capturing EAB beetles. In a recent study in sites with relatively low EAB densities, double-decker traps captured significantly more EAB than green or purple prism traps or green funnel traps, all of which are designed to be suspended from a branch in the canopy of ash trees. A greater percentage of double decker traps were positive, i.e., captured at least one EAB, than the prism traps or funnel traps that were hung in ash tree canopies.

In a large scale study, three artificial trap designs as well as girdled ash trees were deployed systematically, 10-20 m apart, across a newly infested 16 ha forested area with a very low density of EAB25. Artificial trap designs tested included purple prisms baited with Manuka oil and suspended &#62;3 m high from a branch in the canopy of ash trees, 3 m tall DD traps with two green panels, and DD traps with two purple panels supported by t-posts. Double-decker tra...

Watch this Scientific Journal Video about Building Double-decker Traps for Early Detection of Emerald Ash Borer at JoVE.com

Building Double-decker Traps for Early Detection of Emerald Ash Borer

Effective traps to attract and capture the emerald ash borer (EAB) are a key element of detecting and managing this invasive pest. Double-decker traps, placed in full sun near ash trees, incorporate visual and olfactory cues and were more likely to capture EAB than other trap designs in field trials.

Emerald ash borer (EAB) (Agrilus planipennis Fairmaire), the most destructive forest insect to have invaded North America, has killed hundreds of millions ...

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7.8K Views.  Michigan State University. Effective traps to attract and capture the emerald ash borer (EAB) are a key element of detecting and managing this invasive pest. Double-decker traps, placed in full sun near ash trees, incorporate visual and olfactory cues and were more likely to capture EAB than other trap designs in field trials.

Video: Building Double-decker Traps for Early Detection of Emerald Ash Borer

Technique for Studying Arthropod and Microbial Communities within Tree Tissues

Phloem tissues of pine are habitats for many thousands of organisms. Arthropods and microbes use phloem and cambium tissues to seek mates, lay eggs, rear young, feed, or hide from natural enemies or harsh environmental conditions outside of the tree. Organisms that persist within the phloem habitat are difficult to observe given their location under bark. We provide a technique to preserve intact phloem and prepare it for experimentation with invertebrates and microorganisms. The apparatus is called a &#8216;phloem sandwich&#8217; and allows for the introduction and observation of arthropods, microbes, and other organisms. This technique has resulted in a better understanding of the feeding behaviors, life-history traits, reproduction, development, and interactions of organisms within tree phloem. The strengths of this technique include the use of inexpensive materials, variability in sandwich size, flexibility to re-open the sandwich or introduce multiple organisms through drilled holes, and the preservation and maintenance of phloem integrity. The phloem sandwich is an excellent educational tool for scientific discovery in both K-12 science courses and university research laboratories.

We provide a technique to preserve intact tree phloem and prepare it for observation. We create an apparatus called a phloem sandwich that allows for the introduction and observation of arthropods, microbes, and other organisms that inhabit phloem tissues.

Phloem tissues of pine are habitats for many thousands of organisms.  Arthropods and microbes use phloem and cambium tissues to seek mates, lay eggs, rear ...

A Technique to Screen American Beech for Resistance to the Beech Scale Insect (Cryptococcus fagisuga Lind.)

Beech bark disease (BBD) results in high levels of initial mortality, leaving behind survivor trees that are greatly weakened and deformed.&#160;The disease is initiated by feeding activities of the invasive beech scale insect, Cryptococcus fagisuga, which creates entry points for infection by one of the Neonectria species of fungus.&#160;Without scale infestation, there is little opportunity for fungal infection. Using scale eggs to artificially infest healthy trees in heavily BBD impacted stands demonstrated that these trees were resistant to the scale insect portion of the disease complex1.&#160;Here we present a protocol that we have developed, based on the artificial infestation technique by Houston2, which can be used to screen for scale-resistant trees in the field and in smaller potted seedlings and grafts.&#160;The identification of scale-resistant trees is an important component of management of BBD through tree improvement programs and silvicultural manipulation.

A Technique to Screen American Beech for Resistance to the Beech Scale Insect Cryptococcus fagisuga Lind.

Beech bark disease is initiated by feeding activities of the beech scale insect that create fungal entry points in the bark.&#160;Trees that are resistant to the scale insect are also disease resistant.&#160;Here we present the protocol we have developed to screen individual beech trees for beech scale resistance.

Beech bark disease (BBD) results in high levels of initial mortality, leaving behind survivor trees that are greatly weakened and deformed.&#160;The ...

Soil Sampling and Isolation of Entomopathogenic Nematodes (Steinernematidae, Heterorhabditidae)

Entomopathogenic nematodes (a.k.a. EPN) represent a group of soil-inhabiting nematodes that parasitize a wide range of insects. These nematodes belong to two families: Steinernematidae and Heterorhabditidae. Until now, more than 70 species have been described in the Steinernematidae and there are about 20 species in the Heterorhabditidae. The nematodes have a mutualistic partnership with Enterobacteriaceae bacteria and together they act as a potent insecticidal complex that kills a wide range of insect species.
Herein, we focus on the most common techniques considered for collecting EPN from soil. The second part of this presentation focuses on the insect-baiting technique, a widely used approach for the isolation of EPN from soil samples, and the modified White trap technique which is used for the recovery of these nematodes from infected insects. These methods and techniques are key steps for the successful establishment of EPN cultures in the laboratory and also form the basis for other bioassays that consider these nematodes as model organisms for research in other biological disciplines. The techniques shown in this presentation correspond to those performed and/or designed by members of S. P. Stock laboratory as well as those described by various authors.

Soil Sampling and Isolation of Entomopathogenic Nematodes Steinernematidae, Heterorhabditidae

Entomopathogenic nematodes are soil-inhabiting roundworms that parasitize a wide range of insects. We demonstrate sampling methods used for the isolation of these nematodes from the soil using two techniques: the insect baiting and the modified White trap, for the recovery of nematodes from soil samples and infected insect cadavers, respectively.

Entomopathogenic nematodes (a.k.a. EPN) represent a group of soil-inhabiting nematodes that parasitize a wide range of insects. These nematodes belong to ...

A Push-pull Protocol to Reduce Colonization of Bird Nest Boxes by Honey Bees

Introduction of the invasive Africanized honey bee (AHB) into the Neotropics is a serious problem for many cavity nesting birds, specifically parrots. These bees select cavities that are suitable nest sites for birds, resulting in competition. The difficulty of removing bees and their defensive behavior makes a prevention protocol necessary. Here, we describe a push-pull integrated pest management protocol to deter bees from inhabiting bird boxes by applying a bird safe insecticide, permethrin, to repel bees from nest boxes, while simultaneously attracting them to pheromone-baited swarm traps. Shown here is an example experiment using Barn Owl nest boxes. This protocol successfully reduced colonization of Barn Owl nest boxes by Africanized honey bees. This protocol is flexible, allowing adjustments to accommodate a wide range of bird species and habitats. This protocol could benefit conservation efforts where AHB are located.

Preventing colonization of bird nest boxes by invasive Africanized honey bees is important for conservation efforts of nest-site limited birds. We provide an integrated pest management approach to &#34;push&#34; bees away from nest boxes with a repellant insecticide, permethrin, and &#34;pull&#34; them toward pheromone baited swarm traps.

Introduction of the invasive Africanized honey bee (AHB) into the Neotropics is a serious problem for many cavity nesting birds, specifically parrots. ...

The Hawaii Protocol for Scientific Monitoring of Coffee Berry Borer: a Model for Coffee Agroecosystems Worldwide

Coffee berry borer (CBB) is the most devastating insect pest for coffee crops worldwide. We developed a scientific monitoring protocol that is aimed at capturing and quantifying the dynamics and impact of this invasive insect pest as well as the development of its host plant across a heterogeneous landscape. The cornerstone of this comprehensive monitoring system is timely georeferenced data collection on CBB movement, coffee berry infestation, mortality by the fungus Beauveria bassiana, and coffee plant phenology via a mobile electronic data recording application. This electronic data collection system allows field records to be georeferenced through built-in global positioning systems, and is backed by a network of weather stations and records of farm management practices. Comprehensive monitoring of CBB and host plant dynamics is an essential part of an area-wide project in Hawaii to aggregate landscape-level data for research to improve management practices. Coffee agroecosystems in other parts of the world that experience highly variable environmental and socioeconomic factors will also benefit from implementing this protocol, in that it will drive the development of customized integrated pest management (IPM) to manage CBB populations.

Comprehensive monitoring of coffee berry borer and host plant dynamics is essential for aggregating landscape-level data to improve management of this invasive pest. Here, we present a protocol for scientific monitoring of coffee berry borer movement, infestation, mortality, coffee plant phenology, weather, and farm management via a mobile electronic data recording application.

Coffee berry borer (CBB) is the most devastating insect pest for coffee crops worldwide. We developed a scientific monitoring protocol that is aimed at ...

A Precise and Autonomous System for the Detection of Insect Emergence Patterns

Existing systems to measure insect emergence patterns have limitations; they are only partially automated and are limited in the maximum number of emerging insects they can detect. In order to obtain precise measurement of insect emergence, it is necessary for systems to be semi-automated and able to measure large numbers of emerging insects. We addressed these issues by designing and building a system that is automated and can measure emergence of up to 1200 insects. We modified the existing &#34;falling-ball&#34; system using Arduino microcontrollers to automate data collection and expand the sample size through multiple data channels. Multiple data channels enable the user to not only increase their sample size, but also allows for multiple treatments to be run simultaneously in a single experiment. Furthermore, we created an R script to automatically visualize the data as a bubble plot, while also calculating the median day and time of emergence. The current system was designed using 3D printing so the user can modify the system to be adjusted for different species of insects. The goal of this protocol is to investigate important questions in chronobiology and stress physiology, using this precise and automated system to measure insect emergence patterns.

Measurement of insect emergence patterns requires precision. Existing systems are only semi-automated and sample size is limited. We addressed these issues by designing a system using microcontrollers to precisely measure the time of emergence of large numbers of emerging insects.

Existing systems to measure insect emergence patterns have limitations; they are only partially automated and are limited in the maximum number of ...

A Method for Quantifying Foliage-Dwelling Arthropods

Terrestrial arthropods play an important role in our environment. Quantifying arthropods in a way that allows for a precise index or estimates of density requires a method with high detection probability and a known sampling area. While most described methods provide a qualitative or semi-quantitative estimate adequate for describing species presence, richness, and diversity, few provide an adequately consistent detection probability and known or consistent sampling areas to provide an index or estimate with adequate precision to detect differences in abundance across environmental, spatial, or temporal variables. We describe how to quantify leaf-dwelling arthropods by sealing the leaves and end of branches in a bag, clipping and freezing the bagged material, and rinsing the previously frozen material in water to separate arthropods from the substrate and quantify them. As we demonstrate, this method can be used at a landscape scale to quantify leaf-dwelling arthropods with adequate precision to test for and describe how spatial, temporal, environmental, and ecological variables influence arthropod richness and abundance. This method allowed us to detect differences in density, richness, and diversity of leaf-dwelling arthropods among 5 genera of trees commonly found in southeastern deciduous forests.

We describe how to quantify leaf dwelling arthropods by sealing the leaves and end of branches in a bag, clipping and freezing the bagged material, and rinsing the previously frozen material in water to separate arthropods from the substrate for quantification.

Terrestrial arthropods play an important role in our environment. Quantifying arthropods in a way that allows for a precise index or estimates of density ...

Quantifying Corticolous Arthropods Using Sticky Traps

Terrestrial arthropods play an important role in our environment. Quantifying arthropods in a way that allows for a precise index or estimate of density requires a method with high detection probability and a consistent sampling area. We used manufactured sticky traps to compare abundance, total length (a surrogate for biomass), richness, and Shannon diversity of corticolous arthropods among the boles of 5 tree species. Efficacy of this method was adequate to detect variation in corticolous arthropods among tree species and provide a standard error of the mean that was &#60;20% of the mean for all estimates with sample sizes from 7 to 15 individual trees of each species. Our results indicate, even with these moderate sample sizes, the level of precision of arthropod community metrics produced with this approach is adequate to address most ecological questions regarding temporal and spatial variation in corticolous arthropods. Results from this method differ from other quantitative approaches such as chemical knockdown, visual inspection, and funnel traps in that they provide an indication of corticolous arthropod activity over a relatively long-term, better including temporary bole residents, flying arthropods that temporarily land on the tree bole and crawling arthropods that use the tree bole as a travel route from the ground to higher forest foliage. Furthermore, we believe that commercially manufactured sticky traps provide more precise estimates and are logistically simpler than the previously described method of directly applying a sticky material to tree bark or applying a sticky material to tape or other type of backing and applying that to the tree bark.

We describe a semi-quantitative approach of measuring characteristics of corticolous (bark-dwelling) arthropod communities. We placed commercially manufactured sticky traps on tree boles to estimate abundance, total length (a surrogate to biomass), richness, and Shannon diversity for comparison among tree species.

Terrestrial arthropods play an important role in our environment. Quantifying arthropods in a way that allows for a precise index or estimate of density ...

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.

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 ...

Analysis of Gene Expression in Emerald Ash Borer (Agrilus planipennis) Using Quantitative Real Time-PCR

Emerald ash borer (EAB, Agrilus planipennis) is an exotic invasive pest, which has killed millions of ash trees (Fraxinus spp) in North America. EAB continues to spread rapidly and attacks ash trees of different ages, from saplings to mature trees. However, to date very little or no molecular knowledge exists for EAB. We are interested in deciphering the molecular-based physiological processes at the tissue level that aid EAB in successful colonization of ash trees. In this report we show the effective use of quantitative real-time PCR (qRT-PCR) to ascertain mRNA levels in different larval tissues (including midgut, fat bodies and cuticle) and different developmental stages (including 1st-, 2nd-, 3rd-, 4th-instars, prepupae and adults) of EAB. As an example, a peritrophin gene (herein named, AP-PERI1) is exemplified as the gene of interest and a ribosomal protein (AP-RP1) as the internal control. Peritrophins are important components of the peritrophic membrane/matrix (PM), which is the lining of the insect gut. The PM has diverse functions including digestion and mechanical protection to the midgut epithelium.

Analysis of Gene Expression in Emerald Ash Borer Agrilus planipennis Using Quantitative Real Time-PCR

Quantitative real-time PCR (qRT-PCR) is an effective tool to diagnose mRNA levels in different insect tissues and developmental stages. In this report we show the use of qRT-PCR to ascertain mRNA levels in different larval tissues and developmental stages of the invasive insect species, emerald ash borer.

Emerald ash borer (EAB, Agrilus planipennis) is an exotic invasive pest, which has killed millions of ash trees (Fraxinus spp) in North America. EAB ...