Progress 09/01/18 to 04/30/21

Outputs
Target Audience:For the duration of the Tick Surveillance and Mass Capture Trap Phase I project, the target audiences were any populations affected by tick infestation and at risk of tick-transmitted diseases, and those looking for alternatives to acaricides, with a focus on groups who wish to monitor and trap ticks that affect livestock. Our Phase I efforts were directed toward the research and professional land management/pest control communities affecting agricultural livestock. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest?We are preparing a manuscript for publication in a peer-reviewed journal and have submitted an abstract for a 10-min talk at the forthcoming 2021 Annual Meeting of the Entomological Society of America. Because the culmination of the project will result in a commercial product, we have not yet engaged in extension outreach specifically directed at the user community. In our Phase I efforts, we worked with research personnel at the USDA-ARS facility in Pullman, WA/Moscow, ID and with US Navy entomologists at the US Marine Corps base at Camp Lejeune, NC. Their input has been valuable in shaping the prototype device's development so that it will be ready for use by our target audience following efficacy studies. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? During Phase I we developed a commercially-manufacturable prototype tick trap based on the Yans Modular Insect Trap. Our trap is an economical alternative that can be mass-produced rather than being hand-built, and uses an adhesive card that is both quick to deploy and produced at minimal cost on standard equipment. The prototype trap is as effective as the hand-built traps in the field, with an optimized shape and a shelf-stable CO2 attractant that does not differ significantly from dry ice in laboratory studies. Aim 1: Developing a Tick-Targeted Adhesive We formulated an adhesive that is both as effective as carpet tape and compatible with testing captured specimens for pathogens, as it does not leave enough residue behind to interfere with sampling for genetic analyses. 1) Tick capture efficacy Activities: Bioassays to evaluate ability of adhesives to capture ticks Data: Capture rates for some or all life stages of 4 species of ticks (Amblyomma americanum, Dermacentor andersoni, Rhipicephalus microplus, R. sanguineus), genders separated for adults, for up to 10 adhesives Results: Our favored adhesive captured 83% of all adult ticks assayed (n=388), with 63% of those on their dorsum; this was not significantly different from carpet tape, 89% captured (n=201), 65% dorsally. Outcomes: Any glue that captured adults without repelling them would also be strong enough to capture juveniles. Several of the adhesives had repellent effects toward ticks; this opens up a potential new avenue for development of products that have the potential to protect vertebrate hosts from tick bites. 2) Adhesive retention assay Activities: Adhesive bioassays to evaluate adhesive retention on captured ticks; consultation with experts regarding glue retention/tick genetic analyses Data: Whether or not adhesives damaged/destroyed ticks pulled off Results: Glues did not leave detectable amounts of residue on captured ticks Outcomes: Based on advice from tick experts at USDA-ARS facilities in Pullman, WA/Moscow, ID, we abandoned these assays. Any traces of adhesives would either be dissolved or discarded during the DNA extraction preparation process with other impurities. Aim 2: Trap Design for Manufacturability, Durability, Efficacy, and Capacity Our prototype trap has a curved base, which results in a gradually-decreasing angle as it approaches the glue card, based on iterative laboratory testing. Our device is simple to use and deploy, even for untrained personnel, and can form the basis of a non-pesticidal mass-trapping tick control option that could potentially eliminate ticks from a given area when used with the proper protocols. 1) Device size Activities: Analysis of existing products for unit box dimensions, cost per part, and other distribution considerations (e.g. shelf size limitations) Data: Desired range of overall product dimensions Results: Scalable prototype device Outcomes: Based on our observations of tick behavior, the overall device dimensions are less critical to tick captures than the device shape, as ticks will freely climb up most surfaces given enough time. This allows us to be flexible with the shape and customize it to fit with our existing product line, regarding unit boxes, case boxes, printing, etc. 2) Device shape Activities: Ramp angle testing using custom arenas and adhesive cards Data: The optimal range of angles for capturing ticks of 4 species (A. americanum, D. andersoni, R. microplus, R. sanguineus) Results: 98.9% of all ticks captured (n=647) at angles of 9.5°-17° where the ramp approached the adhesive Outcomes: Ticks tightly wedged themselves between the ramp and the card. The gradual sloping allows for ticks of different life stages and sizes to be caught. 3) Manufacturability Activities: CAD-design and 3D-printing of testing arenas and prototype traps; vacuum former molding Data: Whether or not a design would be manufacturable by a given method Results: Prototype single-part device that can be mass-produced by either vacuum-forming or injection molding Outcomes: As long as the device is designed with an appropriate draft angle and without undercuts, it can be vacuum-formed or injection molded, and scaled up/down in size. The simple geometry parameters chosen will allow a low-cost mold tool and low-cost final product. Aim 3: Evaluation of Alternative Attractants 1) Shelf-stable CO2-producing alternatives Activities: Creation of carbohydrate-yeast blends with 4 strains of commercially-available yeasts and 3 carbohydrates in multiple ratios; gravimetric analyses of all blends Data: Instantaneous release rates over multiple days for all blends Results: The ideal ratio for CO2 production from 0-48 h is 1 part yeast to 10 parts carbohydrate,; however, the ratio of 1 part yeast to 20 parts carbohydrate performed optimally from 24-100+ h. Outcomes: A shelf-stable blend that produces relatively high amounts of CO2 over approximately 72 h. Yeast-carbohydrate formulations produce an array of compounds in addition to CO2, many of which are known to attract mosquitoes. These compounds may provide additional effects on different tick species. Shelf stability is comparable to commercially available yeasts found in grocery stores for baking. 2) Tick bioassays Activities: Tick bioassays in Y-tube olfactometer with best-performing biogenic CO2 blends, CO2 produced by dry ice Data: Tick choice data (treatment versus null) for adult/juvenile A. americanum, D. andersoni Results: Selection of biogenic CO2 blend for 72-h duration; candidate yeast-carbohydrate blends for longer duration testing in Phase II Outcomes: CO2 acts as a stimulant that initiates tick movement/questing, not directional attraction. High CO2 levels from dry ice incapacitated ticks, while low levels initiated activity but did not necessarily result in a choice; similar observations were made with biogenic blends. Tick activity increased compared to a null-null test, but they did not necessarily make a directed choice. Aim 4: Field Testing of Optimized Trap 1) Comparison to original trap Activities: Field tests in eastern Washington, North Carolina, and Oklahoma Data: A. americanum capture data from NC and OK for the prototype trap compared to the hand-built original and dry-ice baited sheets. No data from WA due to low local populations of ticks. Results: Overall captures were not significantly different between the Yans trap and the prototype trap in both NC and OK. In OK, none of the monitoring methods were significantly different from the others (F = 0.9999, df = 2, 13, P = 0.9056). In NC, the Yans trap caught more adult females and males than either the prototype or the white sheet, although the prototype was superior to the white sheet for males (F = 8.2798, df = 2,33, P = 0.0012 for females; F = 16.6908, df = 2, 33, P < 0.0001 for males). This order was reversed for nymphs, with the prototype catching the highest numbers, and significantly more than the white sheets (F = 4.0875, df = 2, 13, P = 0.0259). For all life stages combined in NC, the prototype and the Yans trap performed equally, and both caught 3.6x more ticks than the white sheets (F = 6.3957, df = 2, 13, P = 0.0045). Outcomes: The superiority over white sheets and the high numbers of ticks captured in both Yans traps and prototypes during a 4-h period (>40 in OK and >70 in NC) are very encouraging. They suggest that for ambulatory ticks the traps could become effective monitoring tools. The catches suggest that the trap should be evaluated for mass-trapping and population reduction in limited areas. The results indicate that improvements could be made to the BanfieldBio prototype trap so that it catches more adults without sacrificing its excellent performance in capturing nymphs, which we will pursue in Phase II, along with targeting ticks of other species that may be less ambulatory than A. americanum and/or active at a different time of day, like Ixodes spp.

Publications

• Type: Journal Articles Status: Other Year Published: 2021 Citation: Yans, M.W., A.S. Branca, N.G. Hahn, G. Kemble, S.E. Crawley, K.R. Hobson, M.G. Banfield and J.H. Borden. 2021. Development of a simple trap that captures ticks (Acari) on their dorsal surface. Manuscript in preparation for the Journal of Medical Entomology.
• Type: Conference Papers and Presentations Status: Submitted Year Published: 2021 Citation: Yans, M.W., A.S. Branca, N.G. Hahn, S.E. Crawley, K.R. Hobson, M.G. Banfield and J.H. Borden. 2021. Development of a simple trap that captures ticks (Acari) on their dorsal surface. Abstract submitted for 10-min presentation at 2021 Annual Meeting of the Entomological Society of America.

Progress 09/01/18 to 08/31/19

Outputs
Target Audience:The overall target audience for the entire Tick Surveillance and Mass Capture Trap project remains any populations affected by tick infestation and at risk of tick-transmitted diseases, and those looking for alternatives to acaricides. Our Phase I efforts to date have been directed toward the research and professional land management/pest control communities, i.e. individuals or groups interested in monitoring areas for tick populations. Changes/Problems:We encountered two major problems that caused delays in the experimental procedures. The first problem was the government shutdown, which impacted our supply of ticks for laboratory studies. Not only were we unable to order ticks for the duration of the shutdown from the USDA-ARS facility in Pullman, WA/Moscow, ID, we also had to wait for the colony to be rejuvenated following the shutdown. While we were able to get some ticks from another source (BEI Resources), it took some time to be approved for receipt of their shipments, and they were supplying us with a different tick species than the USDA-ARS facility. Thus, we were unable to complete all of our planned laboratory testing with sufficient replication and the species that we originally intended for testing. The second problem is the original term of the award itself, 09/01/2018 - 04/30/2019. Our project was written to culminate in field testing of our laboratory-proven prototype, but the field season for ticks does not fall within the term of the award. In the locations available to us for testing, the field season does not begin until May at the earliest. Knowing this, we requested a no-cost extension, and intend to finish our final testing with field populations of ticks within the next few months. What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest?Because the culmination of the project will result in a commercial product, we have not engaged in any outreach specifically directed at our target audience. In our Phase I efforts to date, we have been working with research personnel at the USDA-ARS facility in Pullman, WA/Moscow, ID and with US Navy entomologists at the US Marine Corps base at Camp Lejeune, NC. Their input has been valuable in shaping the prototype device's development so that it will be ready for use by our target audience following efficacy studies. What do you plan to do during the next reporting period to accomplish the goals?Aim 1: Developing a Tick-Targeted Adhesive 3) Stability in environmental conditions Testing the stability of adhesives under differing environmental conditions was delayed primarily because of low tick populations in USDA-ARS collaborator-owned colonies from which our live ticks were sourced in addition to the government shutdown limiting the availability of ticks for testing from CDC sources. This testing will be done with the next batches of ticks we receive. However, several of the candidate adhesives are already used in other SpringStar traps and we will be able to use information already on hand on their stability under harsh environmental conditions. Aim 4: Field Testing of Optimized Trap 1) Comparison to original 2) Assessing trap longevity Because the field season during which ticks are active and in high populations did not fall within the original grant period (9/1/2018 - 4/30/2019), we will be doing both small-scale field experiments as laid out in the application/project initiation within the next few months. Exact timing of our field studies will depend on tick population levels and seasonality at our study sites.

Impacts
What was accomplished under these goals? During the previous project period we made significant progress toward the development of a commercial tick trap based on the Yans Modular Insect Trap. Our trap is an economical alternative that can be mass-produced rather than being hand-built, and uses an adhesive card that is both quick to deploy and produced at minimal cost on our existing equipment, compared to carpet tape. To date, the prototype trap is as effective as the original hand-built traps, with an optimized shape and a shelf-stable CO2 attractant that does not differ significantly from dry ice in laboratory studies. Ticks transmit a wide range of human and animal pathogens; coupled with their range expansion and invasions of new species, this calls for the development of effective traps for monitoring/control. Ticks reduce animal product yields both through their biting and feeding, and the diseases they vector. Detrimental effects resulting from tick bites and tick-transmitted diseases can include serious reductions in weight gains and milk production; lower market value of hides, fur, or wool; and death in extreme cases. Treatments for controlling ticks on livestock are costly and ticks have exhibited resistance to many commonly-used acaricides. Thus, an effective budget-friendly tool for monitoring and controlling tick populations is warranted. Aim 1: Developing a Tick-Targeted Adhesive We have a favored adhesive that is both as effective as carpet tape and compatible with testing captured specimens for pathogens, as it does not leave enough residue behind to interfere with sampling for genetic analyses. 1) Tick capture efficacy Activities: Tick bioassays to evaluate ability of adhesives to capture ticks dorsally Data: Capture rates for some or all life stages of 3 species of ticks (Amblyomma americanum, Dermacentor andersoni, Rhipicephalus sanguineus), genders separated for adults, for up to 10 different adhesives Results: Our favored adhesive captured 52% of assayed ticks on their dorsum. This was not significantly different from the percent of ticks (56%) captured on carpet tape. Outcomes: Any glue that captured adults without repelling them would also be strong enough to capture juveniles. Several of the adhesives had repellent effects toward ticks; this opens up a potential new avenue for testing. 2) Adhesive retention assay Activities: Adhesive bioassays to evaluate amount of adhesive retained by captured ticks; consultation with experts regarding glue retention/tick genetic analyses Data: Whether or not adhesives damaged/destroyed ticks pulled off Results: Best-performing glues did not leave detectable amounts of residue on captured ticks Outcomes: Based on advice from tick experts at USDA-ARS facilities in Pullman, WA/Moscow, ID, we abandoned these assays. Any tick samples would be exposed to various solvents during the DNA extraction procedure, and any traces of adhesives would either be dissolved or discarded during the sample preparation process with any other impurities. 3) Stability in environmental conditions Activities: Aging of adhesive cards in varied environmental conditions Data/Results/Outcomes: Not tested yet, see Project Changes Aim 2: Trap Design for Manufacturability, Durability, Efficacy, and Capacity Our prototype trap has a ramp angle that gradually decreases as it approaches the glue card. This design was based on laboratory testing with all tick life stages and feedback from our US Navy collaborator who tested several iterations of prototypes. Our device is simple to use and deploy, even for untrained personnel, and can form the basis of a non-pesticidal mass-trapping tick control option that could potentially eliminate ticks from a given area when used with the proper protocols. 1) Device size Activities: Analysis of existing products for unit box dimensions, cost per part, and other distribution considerations (e.g. shelf size limitations) Data: Desired range of overall product dimensions Results: Scalable prototype device Outcomes: Based on our observations of tick behavior, the overall device dimensions are less critical to tick captures than the device shape, as ticks will freely climb up most surfaces given enough time. This allows us to be flexible with the shape and customize it to fit with our existing product line, regarding unit boxes, case boxes, printing, etc. 2) Device shape Activities: Ramp angle testing using custom arenas/prototype traps and adhesive cards; selection of prototype device ramp angles Data: The optimal range of angles for capturing ticks of 3 species (A. americanum, D. andersoni, R. sanguineus) Results: 99.1% of all ticks tested (n=857) were caught between 9.5°-17° where the ramp approached the adhesive card Outcomes: We found that the angle at which ticks approached the glue card seemed to lead to better captures, as the ticks appeared to wedge themselves in tight enclosed crevices. The gradual sloping allows for ticks of different life stages and sizes to be caught. Larval ticks were observed to be capable of climbing up any angle; however, their climbing ability was limited due to their inability to overcome static electricity when climbing some plastics. 3) Manufacturability Activities: CAD-design and 3D-printing of prototype testing arenas and prototype traps; vacuum former mold testing Data: Whether or not a design would be manufacturable by a given method Results: Prototype design of a single-part device that can be mass-produced by either vacuum-forming or injection molding Outcomes: As with the overall size, the manufacturing process is not critical to the capture of ticks. As long as the device is designed with an appropriate draft angle and without undercuts, it can be vacuum-formed or injection molded, and it can be scaled up or down in size. Aim 3: Evaluation of Alternative Attractants 1) Shelf-stable CO2-producing alternatives Activities: Creation of carbohydrate-yeast blends with four different strains of commercially-available yeasts and three carbohydrates in multiple ratios; gravimetric analyses of biogenic CO2 blends Data: Instantaneous release rates over multiple days for all blends Results: The ideal ratio for CO2 production from 0-48 h is 1 part yeast to 10 parts carbohydrate, with an instantaneous rate of CO2 production described by a linear regression equation of y = -1.01x + 94.22 mL/h; however, the ratio of 1 part yeast to 20 parts carbohydrate performed optimally from 24-100+ hours, with y = -0.23x + 53.99 mL/h. Outcomes: We have a shelf-stable yeast blend that produces relatively high amounts of CO2 over approximately 72 hours. Yeast-carbohydrate formulations are known to produce an array of compounds in addition to CO2, many of which are known to attract mosquitoes. These compounds may provide additional effects on different tick species. 2) Tick bioassays Activities: Tick bioassays in Y-tube olfactometer with best-performing biogenic CO2 blends, CO2 produced by dry ice Data: Tick choice data (treatment versus null) for adult/juvenile A. americanum, D. andersoni Results: Selection of biogenic CO2 blend to use in Aim 4 Outcomes: Our data suggest that CO2 acts as a stimulant that initiates tick movement/questing, not directional attraction. High CO2 levels from dry ice incapacitated ticks, while small amounts initiated activity at the base of the y-tube but did not necessarily result in a choice; similar observations were made with biogenic blends. Tick activity increased compared to a null-null test, but they did not necessarily make a directed choice. Due to the government shut-down and low colony populations, we were not able to acquire sufficient ticks to draw conclusions. Aim 4: Field Testing of Optimized Trap 1) Comparison to original Activities/Data/Results/Outcomes: Not tested yet, see Project Changes 2) Assessing trap longevity Activities/Data/Results/Outcomes: Not tested yet, see Project Changes

Publications

Progress 09/01/18 to 04/30/19

Outputs
Target Audience:The overall target audience for the entire Tick Surveillance and Mass Capture Trap project remains any populations affected by tick infestation and at risk of tick-transmitted diseases, and those looking for alternatives to acaricides. Our Phase I efforts to date have been directed toward the research and professional land management/pest control communities, i.e. individuals or groups interested in monitoring areas for tick populations. Changes/Problems:We encountered two major problems that caused delays in the experimental procedures. The first problem was the government shutdown, which impacted our supply of ticks for laboratory studies. Not only were we unable to order ticks for the duration of the shutdown from the USDA-ARS facility in Pullman, WA/Moscow, ID, we also had to wait for the colony to be rejuvenated following the shutdown. While we were able to get some ticks from another source (BEI Resources), it took some time to be approved for receipt of their shipments, and they were supplying us with a different tick species than the USDA-ARS facility. Thus, we were unable to complete all of our planned laboratory testing with sufficient replication and the species that we originally intended for testing. The second problem is the original term of the award itself, 09/01/2018 - 04/30/2019. Our project was written to culminate in field testing of our laboratory-proven prototype, but the field season for ticks does not fall within the term of the award. In the locations available to us for testing, the field season does not begin until May at the earliest. Knowing this, we requested a no-cost extension, and intend to finish our final testing with field populations of ticks within the next few months. What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest?Because the culmination of the project will result in a commercial product, we have not engaged in any outreach specifically directed at our target audience. In our Phase I efforts to date, we have been working with research personnel at the USDA-ARS facility in Pullman, WA/Moscow, ID and with US Navy entomologists at the US Marine Corps base at Camp Lejeune, NC. Their input has been valuable in shaping the prototype device's development so that it will be ready for use by our target audience following efficacy studies. What do you plan to do during the next reporting period to accomplish the goals?Aim 1: Developing a Tick-Targeted Adhesive 3) Stability in environmental conditions Testing the stability of adhesives under differing environmental conditions was delayed primarily because of low tick populations in USDA-ARS collaborator-owned colonies from which our live ticks were sourced in addition to the government shutdown limiting the availability of ticks for testing from CDC sources. This testing will be done with the next batches of ticks we receive. However, several of the candidate adhesives are already used in other SpringStar traps and we will be able to use information already on hand on their stability under harsh environmental conditions. Aim 4: Field Testing of Optimized Trap 1) Comparison to original 2) Assessing trap longevity Because the field season during which ticks are active and in high populations did not fall within the original grant period (9/1/2018 - 4/30/2019), we will be doing both small-scale field experiments as laid out in the application/project initiation within the next few months. Exact timing of our field studies will depend on tick population levels and seasonality at our study sites.

Impacts
What was accomplished under these goals? During the previous project period we made significant progress toward the development of a commercial tick trap based on the Yans Modular Insect Trap. Our trap is an economical alternative that can be mass-produced rather than being hand-built, and uses an adhesive card that is both quick to deploy and produced at minimal cost on our existing equipment, compared to carpet tape. To date, the prototype trap is as effective as the original hand-built traps, with an optimized shape and a shelf-stable CO2 attractant that does not differ significantly from dry ice in laboratory studies. Ticks transmit a wide range of human and animal pathogens; coupled with their range expansion and invasions of new species, this calls for the development of effective traps for monitoring/control. Ticks reduce animal product yields both through their biting and feeding, and the diseases they vector. Detrimental effects resulting from tick bites and tick-transmitted diseases can include serious reductions in weight gains and milk production; lower market value of hides, fur, or wool; and death in extreme cases. Treatments for controlling ticks on livestock are costly and ticks have exhibited resistance to many commonly-used acaricides. Thus, an effective budget-friendly tool for monitoring and controlling tick populations is warranted. Aim 1: Developing a Tick-Targeted Adhesive We have a favored adhesive that is both as effective as carpet tape and compatible with testing captured specimens for pathogens, as it does not leave enough residue behind to interfere with sampling for genetic analyses. 1) Tick capture efficacy Activities: Tick bioassays to evaluate ability of adhesives to capture ticks dorsally Data: Capture rates for some or all life stages of 3 species of ticks (Amblyomma americanum, Dermacentor andersoni, Rhipicephalus sanguineus), genders separated for adults, for up to 10 different adhesives Results: Our favored adhesive captured 52% of assayed ticks on their dorsum. This was not significantly different from the percent of ticks (56%) captured on carpet tape. Outcomes: Any glue that captured adults without repelling them would also be strong enough to capture juveniles. Several of the adhesives had repellent effects toward ticks; this opens up a potential new avenue for testing. 2) Adhesive retention assay Activities: Adhesive bioassays to evaluate amount of adhesive retained by captured ticks; consultation with experts regarding glue retention/tick genetic analyses Data: Whether or not adhesives damaged/destroyed ticks pulled off Results: Best-performing glues did not leave detectable amounts of residue on captured ticks Outcomes: Based on advice from tick experts at USDA-ARS facilities in Pullman, WA/Moscow, ID, we abandoned these assays. Any tick samples would be exposed to various solvents during the DNA extraction procedure, and any traces of adhesives would either be dissolved or discarded during the sample preparation process with any other impurities. 3) Stability in environmental conditions Activities: Aging of adhesive cards in varied environmental conditions Data/Results/Outcomes: Not tested yet, see Project Changes Aim 2: Trap Design for Manufacturability, Durability, Efficacy, and Capacity Our prototype trap has a ramp angle that gradually decreases as it approaches the glue card. This design was based on laboratory testing with all tick life stages and feedback from our US Navy collaborator who tested several iterations of prototypes. Our device is simple to use and deploy, even for untrained personnel, and can form the basis of a non-pesticidal mass-trapping tick control option that could potentially eliminate ticks from a given area when used with the proper protocols. 1) Device size Activities: Analysis of existing products for unit box dimensions, cost per part, and other distribution considerations (e.g. shelf size limitations) Data: Desired range of overall product dimensions Results: Scalable prototype device Outcomes: Based on our observations of tick behavior, the overall device dimensions are less critical to tick captures than the device shape, as ticks will freely climb up most surfaces given enough time. This allows us to be flexible with the shape and customize it to fit with our existing product line, regarding unit boxes, case boxes, printing, etc. 2) Device shape Activities: Ramp angle testing using custom arenas/prototype traps and adhesive cards; selection of prototype device ramp angles Data: The optimal range of angles for capturing ticks of 3 species (A. americanum, D. andersoni, R. sanguineus) Results: 99.1% of all ticks tested (n=857) were caught between 9.5°-17° where the ramp approached the adhesive card Outcomes: We found that the angle at which ticks approached the glue card seemed to lead to better captures, as the ticks appeared to wedge themselves in tight enclosed crevices. The gradual sloping allows for ticks of different life stages and sizes to be caught. Larval ticks were observed to be capable of climbing up any angle; however, their climbing ability was limited due to their inability to overcome static electricity when climbing some plastics. 3) Manufacturability Activities: CAD-design and 3D-printing of prototype testing arenas and prototype traps; vacuum former mold testing Data: Whether or not a design would be manufacturable by a given method Results: Prototype design of a single-part device that can be mass-produced by either vacuum-forming or injection molding Outcomes: As with the overall size, the manufacturing process is not critical to the capture of ticks. As long as the device is designed with an appropriate draft angle and without undercuts, it can be vacuum-formed or injection molded, and it can be scaled up or down in size. Aim 3: Evaluation of Alternative Attractants 1) Shelf-stable CO2-producing alternatives Activities: Creation of carbohydrate-yeast blends with four different strains of commercially-available yeasts and three carbohydrates in multiple ratios; gravimetric analyses of biogenic CO2 blends Data: Instantaneous release rates over multiple days for all blends Results: The ideal ratio for CO2 production from 0-48 h is 1 part yeast to 10 parts carbohydrate, with an instantaneous rate of CO2 production described by a linear regression equation of y = -1.01x + 94.22 mL/h; however, the ratio of 1 part yeast to 20 parts carbohydrate performed optimally from 24-100+ hours, with y = -0.23x + 53.99 mL/h. Outcomes: We have a shelf-stable yeast blend that produces relatively high amounts of CO2 over approximately 72 hours. Yeast-carbohydrate formulations are known to produce an array of compounds in addition to CO2, many of which are known to attract mosquitoes. These compounds may provide additional effects on different tick species. 2) Tick bioassays Activities: Tick bioassays in Y-tube olfactometer with best-performing biogenic CO2 blends, CO2 produced by dry ice Data: Tick choice data (treatment versus null) for adult/juvenile A. americanum, D. andersoni Results: Selection of biogenic CO2 blend to use in Aim 4 Outcomes: Our data suggest that CO2 acts as a stimulant that initiates tick movement/questing, not directional attraction. High CO2 levels from dry ice incapacitated ticks, while small amounts initiated activity at the base of the y-tube but did not necessarily result in a choice; similar observations were made with biogenic blends. Tick activity increased compared to a null-null test, but they did not necessarily make a directed choice. Due to the government shut-down and low colony populations, we were not able to acquire sufficient ticks to draw conclusions. Aim 4: Field Testing of Optimized Trap 1) Comparison to original Activities/Data/Results/Outcomes: Not tested yet, see Project Changes 2) Assessing trap longevity Activities/Data/Results/Outcomes: Not tested yet, see Project Changes

Publications