NE1943: Biology, Ecology & Management of Emerging Disease Vectors

(Multistate Research Project)

Status: Active

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SAES-422 Reports

Annual/Termination Reports:

[01/18/2021] [12/13/2022]

Date of Annual Report: 01/18/2021

Report Information

Annual Meeting Dates: 11/23/2020 - 11/23/2020
Period the Report Covers: 10/01/2019 - 09/30/2020

Participants

Paul Leisnham, University of Maryland; Susan Paskewitz, University of Wisconsin; Allison Gardner, University of Maine; Anna Rosalee Pasternak (student of Subba Palli), University of Kentucky; James Occi (student of Dina Fonseca), Rutgers University; Adela Oliva Chavez, Texas A&M; Emily Reed (student of Michael Reiskind), North Carolina State University; Michael Reskind, North Carolina State University; Erica Kistner Thomas, USDA representative

Brief Summary of Minutes

The annual meeting was organized as an Organized Meeting during the 2020 Entomological Society of America (ESA) Annual Meeting. It consisted of an on-demand session with seven 12-minute seminars and a 1-hr video chat.

The speakers and their seminar titles were as follows:

Dr. Paul Leisnham, University of Maryland: New Insights into the Effects of Tire Leachate on Urban Mosquito Communities

Dr. Susan Paskewitz, University of Wisconsin: Assessments of Vector Control Strategies in the Upper Midwest.

Dr. Allison Gardner, University of Maine: Tick-borne Disease Ecology and Management in Maine.

Anna Rosalee Pasternak, University of Kentucky: Getting Ticky in Kentucky: State-wide Surveillance and Pathogen Testing of Ticks

James Occi, Rutgers University: First Record of Soft Ticks in New Jersey and their Public Health Risk

Dr. Adela S. Oliva Chavez, Texas A&M: Epigenetic Drivers in the Transmission of Tick-borne Diseases in the US.

Emily Reed, North Carolina State University - Population Genetics of an Invasive Mosquito Along an Urban-rural Landscape.

During the 1-hr video chat, the Business Meeting was held. Dr. Leisnham chairs the meeting, first allowing each speaker to introduce themselves and summarize their work. Other attendees then contribute, summarising additional work. Dr. Leisnham begins a discussion by reminding attendees that this project started in October 2019 and will extend to September 2024, building on two prior multistate projects of the same name, which have been very projective and successful. Dr. Leisnham mentions that the preceding multistate project (NE1443) had staged its annual meeting in March to coincide with the annual arbovirus surveillance and mosquito control workshop at the Anastasia Mosquito Control District in St. Augustine, FL. Through email correspondence, project participants decided it was too soon after the establishment of this project to hold this annual meeting in March again; hence, the decision to stage the meeting at the Entomological Society of America's annual meeting. There was then a short discussion on when the next annual meeting should be held, with a recognition that it is always a difficult decision given the conflicting time schedules of project participants. Three society conferences: ESA, the American Society of Tropical Medicine & Hygiene, and the Society of Vector Ecology were identified as arguably the best venues to stage future annual meetings and maximize participant attendance, but that attendance in 2021 would depend heavily on the management of the COVID-19 pandemic. Dr. Leisnham continued the meeting by emphasizing the diversity of the 30 official project participants and their institutions, that we need to contribute to an annual report, and that the project will need to decide on a new Chair. Dr. Leisnham explained that chairs of the preceding projects had served 2-yr appointments and that his 2-yr appointment straddled the end of NE1443 and this project (NE1943). Attendees agreed that 2-yr terms were suitable and there was a consensus to seek a new Chair over the coming months, before the next annual meeting. Last, USDA's representative on this project, Erica Kistner Thomas, introduced herself and briefly described her role. The Meeting adjourned on time after 1 hour.

Accomplishments

Dr. Paul Leisnham and his student, Oswaldo Villena, continued work on tire leachate and mosquito ecology at the University of Maryland. Specifically, they tested the hypothesis that more degraded tires contain greater tire leachate and alter interspecific mosquito competition, producing a condition-specific advantage for the competitively inferior resident, Culex pipiens, by relaxing the effects of competition with the invasive Aedes albopictus. Varying densities of newly hatched Ae. albopictus and Cx. pipiens larvae were added to tires that had been exposed to different ultra-violet (U.V.)-B conditions that mimicked either full-sun, shade, or no UV-B conditions in the field. There were higher competitive effects of Cx. pipiens on the population performance and survival of Ae. albopictus in tires exposed to shade and full-sun U.V. conditions that had higher concentrations of zinc, a marker contaminant of tire leachate. Zinc concentration was also higher in Ae. albopictus than Cx. pipiens indicating greater exposure of the invasive species to tire leachate. These results indicate that tire leachate can affect the outcome of the competition between Ae. albopictus and Cx. pipiens, helping foster the regional persistence of the resident Cx. pipiens. Cx. pipiens is the main vector circulating West Nile virus (WNv) within avian populations in many cities in the northern United States. Ae. albopictus may play an important role in WNv transmission, bridging the virus from avian into human populations. Increased coexistence of these two species resulting from condition-specific competition in tire habitats may increase WNV transmission risk. Research at the Midwest Center of Excellence for Vector-Borne Disease, headed by Dr. Susan Paskewitz of the University of Wisconsin, explored control efforts for the blacklegged tick, Ixodes scapularis, and the northern house mosquito, Cx. pipiens. These two species are responsible for most cases of vector-borne human disease in the upper Midwest and are a focus for this project. Tick control is most often performed at the individual household level.  Using the Tick App and field sampling, researchers confirmed that backyard environments can be important sources of blacklegged ticks in Wisconsin.  Researchers evaluated the efficacy of several low cost, do-it-yourself strategies for homeowners.  Perimeter treatments with a granular formulation of cyahalothrin were highly effective in reducing tick encounters.  Deployment of tick tubes, containing host-targeted nest materials, was less effective although reduction in the density of infected nymphs could still be detected.  Combining tick tubes with removal of invasive vegetation did not increase the effectiveness of treatment.  By comparison, mosquito control for reduction of West Nile virus risk is often done by professional, tax-funded mosquito control operations.  Control of Cx. pipiens focuses on larval control through catch basement treatments and on application of adulticides.  Researchers tested the impacts of these strategies on adult mosquito populations in Chicago and Milwaukee.  While larval control in the basins was good, there was little evidence that this impacted adult populations.  The main impact of application of adulticides was a change in age structure of the population that favored younger mosquitoes. Dr. Allison Gardner and the Vector Biology Lab of the University of Maine completed a study that investigated the impacts of active forest management on the risk of exposure to ticks and tick-borne pathogens across spatial scales, as well as the casual ecological mechanisms underlying observed patterns. Dr. Gardner also introduced an active surveillance citizen science project that seeks to understand the economic, environmental, and production factors that influence private forest landowners’ forest management decision-making processes and the implications for tick-borne disease exposure risk in the landscape. Graduate student Anna Pasternak and her advisor Dr. Subba Reddy Palli of the University of Kentucky conducted Kentucky-wide surveillance of tick distributions at the county level and pathogen testing for detection of Rickettsia rickettsii, Ehrlichia chaffeensis and Borrelia burgdorferi. Preliminary data supports the presence of established lone star tick, Amblyomma americanum, American dog tick, Dermacentor variabilis, and Ixodes scapularis populations infected with Ehrlichia chaffeensis, Rickettsia rickettsia and Borrelia burgdorferi, respectively. These results indicate that Kentuckians are at risk for these bacterial diseases and calls for the implementation of the largescale surveillance program that this study provides. Under the supervision of his advisor Dr. Dina Fonseca from Rutgers University, graduate student James Occi has described the first occurrence of the soft tick Carios kelleyi (Cooley and Kohls) from New Jersey, based on larvae collected from big brown bats, Eptesicus fuscus. Although C. kelleyi is known to occur on bats in at least 29 of the 48 conterminous U.S. states, its ecology of is not well understood, despite reports of this species feeding on humans and its consequent potential as a disease vector. The association of C. kelleyi with bat species that regularly roost in human-made structures, such as attics and barns, and recent isolations from this tick of pathogens capable of infecting humans, companion animals, and livestock underscore the need for further studies of these bat ectoparasites. Dr. Adela Oliva Chavex and her colleagues of Texas A&M University conducted research that sought to determine the role of epigenetics in the vectorial capacity of different tick populations. Researchers investigated the variation in methylation of I. scapularis collected in Minnesota (high-Lyme disease area) and Texas (low-Lyme disease area). Global methylation was assessed by ELISA and bisulfite genome sequencing. The expression of DNA methyltransferases was evaluated by Reverse-Transcriptase (R.T.)-PCR. Preliminary results indicated that the genomes of ticks collected from MN and TX are differentially methylated. Further, bisulfite methylation analysis of TX ticks has identified differential methylation between sexes. These results suggest that similar to bees, DNA methylation plays a role in the life cycle of I. scapularis, one that may influence Lyme disease transmission. Graduate student Emily Reed and her advisor Dr. Michael Reiskind of North Carolina State University completed work exploring the impact of urban landscapes on genetic connectivity of Ae. albopictus. Researchers collected adult mosquitoes across Wake County, North Carolina from 60 locations and built genomic libraries of 336 Ae. albopictus individuals using double digest restriction-site associated DNA sequencing (ddRADseq). Researchers processed sequence data with the STACKS de novo pipeline and analyzed genetic structure, differentiation, and inbreeding between populations with STRUCTURE, DAPC, and popgraph. Using spatial data, researchers then examined how local landscape features correlate with genetic signatures at several spatial scales. This project contributes to a growing body of genetic research that demonstrates how environmental processes affect the genetic structure and evolution of invasive species at fine scales. These studies can improve existing programs for controlling invasive species and will help fill knowledge gaps needed for effective management. Led by Dr. Goudarz Molaei, researchers at the Connecticut Agricultural Experimental Station have worked on four studies. First, they conducted work exploring vector-host interactions of Ae. albopictus in Virginia. Engorged Ae. albopictus were collected from a variety of habitat types using the Centers for Disease Control and Prevention light traps, Biogents Sentinel 2 traps, and modified Reiter gravid traps in southeast Virginia. Sources of blood meals were determined by the analysis of mitochondrial cytochrome b gene sequences amplified in PCR assays. Degrees of Ae. albopictus interactions with vertebrate hosts were quantified, the influence of key socioecological conditions on spatial variability in Ae. albopictus blood feeding was assessed, and temporal differences in blood feeding by season were investigated. Analysis of 961 engorged specimens of Ae. albopictus sampled between 2017-2019 indicated that 96%, 4%, and less than 1% obtained blood meals from mammalian, reptilian, and avian hosts, respectively. Domestic cats were the most frequently identified (50%) hosts for Ae. albopictus followed by Virginia opossums (17%), white-tailed deer (12%), and humans (7%), together representing 86% of all identified blood hosts. A small proportion of blood meals acquired from avian hosts in mixed blood meals suggests that this species may rarely mediate epidemic/epizootic transmission of arboviruses as a bridge vector. Screening of the head and thorax of engorged Ae. albopictus mosquitoes by cell culture and RT-PCR resulted in a single isolate of Potosi virus. Spatial patterns in blood feeding were found to be linked to socioecological conditions and seasonal shifts in Ae. albopictus blood feeding with implications for understanding human biting and disease risk.  In Suffolk Virginia in areas of lower human development, the likelihood of human blood feeding increased as median household income increased and human blood feeding was more likely early in the season (May-June) compared to later (July-October). Understanding of the mosquito-host interactions in nature is vital for evaluating vectorial capacity in mosquitoes and reservoir competency in vertebrate hosts in transmission, maintenance, and amplification of zoonotic agents of human diseases. Results of our study in conjunction with abundance in urban/suburban settings, virus isolations from field-collected mosquitoes, and vector competence of Ae. albopictus, highlight the potential of this species in the transmission of a number of arboviruses such as dengue, chikungunya, and Zika to humans. A manuscript has been submitted to PLoS NTD and is currently under review. Second, Dr. Molaei and his colleagues studied climatic and environmental determinants of the spatial distribution and abundance of I. scapularis (blacklegged tick) and A. americanum (lone star tick). Spatially explicit statistical models were used to link extensive passive tick surveillance data to mean annual temperature and the Wildland-Urban Interface. Blacklegged ticks, endemic in Connecticut, were not associated with mean annual temperature while lone star ticks, recently reported to be established in Connecticut, were positively associated with mean annual temperature. Blacklegged tick submission rates were higher in towns with a higher proportion of land classified as intermix, areas where houses and undeveloped wildland vegetation mix. Lone star tick submission rates were higher in towns with a lower proportion of very low housing density. These findings have implications for tick-borne disease dynamics in the northeastern U.S. as the climate continues to warm and for land use planning to mitigate tick-borne disease risk. A manuscript has been submitted to the CDC Emerging Infections Diseases (EID) and is currently under review. Dr. Molaei identified the first reported established population of Gulf Coast ticks (Amblyomma maculatum) infected with Rickettsia parkeri in Connecticut, representing the northernmost range limit of this medically relevant tick species. This finding highlights the importance of tick surveillance and public health challenges posed by geographic expansion of tick vectors and their pathogens. A manuscript has been submitted to the Journal of Medical Entomology and is currently under review.  Last, Dr. Molaei and his colleagues continued the Connecticut Agricultural Experiment Station (CAES)-Passive Tick Surveillance Program (PTSP) that was established in 1990. This program monitors: 1) tick vector populations for distribution, abundance and range expansion of existing species, 2) introduction of exotic/invasive tick vectors and tick-borne diseases, 3) spatiotemporal dynamics of activity of existing pathogens transmitted by tick vectors to determine the risk of human infection, and 4) conducts investigations on tick biology, ecology, and their roles in disease transmission.  Each year, an average of 3,000 ticks are submitted by the state residents, health departments, and physicians’ offices; however, in recent years the number of submissions has substantially increased, and in some years, it has reached to nearly 6,000. From 10/1/2019 to 9/30/2020, the CAES-PTSP received a total of 3,711 tick submissions. Of these, 3,175 (85.6%) were identified as I. scapularis (blacklegged tick), 369 (9.9%) as D. variabilis (American dog tick), 156 (4.2%) A. americanum (lone star tick), and 11 (0.3%) a few other species. Of the 3,711 total tick submissions, 3,011 (94.8%) engorged nymph and adult female I. scapularis were tested for evidence of infection. Of these, 28.8% (n=867) tested positive for B. burgdorferi, 4.9% (n=149) for Anaplasma phagocytophilum, and 5.4% (n=162) for Babesia microti. Coinfections with B. burgdorferi and A. phagocytophilum were identified in 2.1% (n=63); B. burgdorferi and B. microti in 2.2% (n=66); A. phagocytophilum and B. microti 0.3% (n=9); and with all three pathogens/parasites in 0.2% (n=5). Further research at CAES, under the leadership of Dr. Philip Armstrong, has explored the ecology and control of disease vectors through three projects. The first project continues longstanding work understanding environmental determinants of Ae. albopictus abundance at a northern range limit. Dr. Armstrong ans colleagues constructed linear models to evaluate how trapping methodology, land cover, as well as temperature and precipitation influenced A. albopictus abundance at the northern limits of its range in New York and Connecticut. BGS traps were 2.78 times as efficient as gravid traps and 1.49 times as efficient as CO2-baited CDC light traps. Low and medium-intensity development and low proportions of deciduous cover around the trap site were positively associated with increased abundance, as were minimum winter temperature and March precipitation. The cumulative precipitation within a 28-day time window before the date of collection had a nonlinear relationship with abundance, such that as precipitation increased beyond 70 mm, there was a decrease in abundance. This study shows the impact of landscape and climate variables on A. albopictus distribution at the northern limit of its current range. The second study led by Dr. Armstrong found that successive bloodmeals enhance virus dissemination within mosquitoes and increase transmission potential. Vector competence and the extrinsic incubation period (EIP) are two key entomological parameters used to assess the public health risk posed by arboviruses. These are typically measured by offering mosquitoes an infectious bloodmeal and temporally sampling mosquitoes to determine infection and transmission status. However, this approach does not accurately capture the biology and behavior of many mosquito vectors which refeed frequently (every 2-3 days). The work showed that a second non-infectious bloodmeal significantly shortens the extrinsic incubation period (EIP) of Zika virus (ZIKV) in adult Ae. aegypti by enhancing virus escape from the mosquito midgut. Similarly, a second bloodmeal increased the competence of this species for dengue and chikungunya viruses as well. This effect was also observed for ZIKV in A. albopictus. Bloodmeals induced fissures in the virus-impenetrable basal lamina surrounding the midgut providing a mechanism for enhanced virus escape. Modeling of these findings revealed that a shortened EIP would result in a significant increase in the basic reproductive number, R0. This study helps explain how A. aegypti can sustain an explosive epidemic like ZIKV despite its relatively poor vector competence in single-feed laboratory trials. Last, Dr. Armstrong and colleagues published research that evaluated of novel trapping lures for monitoring exotic and native container-breeding Aedes mosquitoes. During 2018, researchers tested new scent lures, TrapTech Lure-A and Lure-H (Bedoukian Research, Inc.), using BG-Sentinel traps with CO2 in two regions of Connecticut, Stamford and Hamden, against the BG-Lure. Pooled mosquitoes were additionally screened for arbovirus infection.  A total of 47,734 mosquitoes representing 8 genera and 32 species were captured during the study, with the Stamford site deriving on average three times as many mosquitoes per trap, adjusting for effort.  Lure-A and Lure-H outperformed the BG-Lure in terms of overall numbers, diversity evenness, and the proportion of both Ae. japonicus and Aedes triseriatus. There were no significant differences among lures in capturing Ae. albopictus, and in terms of species richness.  Fifty-seven isolates of virus (West Nile, Jamestown Canyon, and La Crosse viruses) were obtained during the study, with no significant difference between trap-lure.  Both novel lures were highlighted as effective potential attractants for use in combination with CO2 in mosquito surveillance.  Dr. Gabe Hamer of Texas A&M has made progress studying the ecology of Aedes aegypti and associated arboviruses in Texas and Mexico and has several related manuscripts published in 2020 or in preparation for submission (see publications). These include evaluating multiple control tools, including a Autocidal Gravid Ovitrap Intervention which has a manuscript in review.  The intervention using the Autodissemination Stations with pyriproxyfen was originally planned for the summer of 2020 but was postponed to 2021 due to COVID.  Another manuscript is in preparation looking at the unique human dimensions of community engagement in South Texas. Dr. Stephen Dobson at the University of Kentucky has been focused on developing improved monitoring tools that combine internet-connected databases and quality control with the multiple trap types being used by mosquito abatement districts, e.g., B.G. traps, CDC traps, ovitraps, etc. The data is updated in real time as collections are made and then presented as a ‘Dashboard’ for managers and supervisors, who can then generate standard and customize report summaries for stakeholders. Furthermore, the sample collections can be tracked through subsequent assays and tests, e.g., PCR and virus assays, to avoid tracking errors. Dr. Zhijian Jake Tu at Virginia Polytechnic Institute and State University have improved the characterization of the male-determining locus (M) in the dengue and Zika vector Ae. aegypti. The key improvement is the determination of the content of the ~160 Kilobase gap in the M-locus. The red-eye mutation within a 5 Mb region that is linked to the M locus was mapped. This is a naturally occurring mutation that gives predominantly wild-type males (only ~2% wild-type individuals are females while most of the females are red-eyed) in progeny. A few candidate genes within this region have been identified. Dr. Bruce Noden from Oklahoma State University is conducting work describing the regional spread of Culex coronator, an invasive mosquito species which has spread throughout the southern U.S. in the last 20 years. So far, they have reported Cx. coronator in 16 of Oklahoma’s 77 counties with expansion throughout the state.

Publications

Armstrong PM, Ehrlich HY, Magalhaes T, Miller MR, Conway PJ, Bransfield A, Misencik MJ, Gloria-Soria A, Warren JL, Andreadis TG, Shepard JJ, Foy BD, Pitzer VE, Brackney DE (2020) Successive blood meals enhance virus dissemination within mosquitoes and increase transmission potential. Nat Microbiol 5(2):239-247. Aryan, A., Anderson M., Biedler J.K., Qi, Y., Overcas J. M., Naumenko A., Sharakhova M.A., Mao C., Adelman Z.A. and Tu Z. 2020. Nix alone is sufficient to convert female Aedes aegypti into fertile males and myo-sex is needed for male flight. Proc Natl Acad Sci U S A. 117 (30), 17702-17709. Compton A, Liang J, Chen C, Lukyanchikova V, Qi Y, Potters M, Settlage R, Miller D, Deschamps S, Mao C, Llaca V, Sharakhov IV, Tu Z. 2020. The beginning of the end: a chromosomal assembly of the New World malaria mosquito ends with a novel telomere. G3 Genes|Genomes|Genetics doi: 10.1534/g3.120.401654<a href="https://www.biorxiv.org/content/10.1101/2020.04.17.047084v2">,</a> 10, 3811-3819 Compton, A., Sharakhov, IV and Tu, Z. 2020. Recent Advances and Future Perspectives in Vector-omics. Current Opinion in Insect Science. https://doi.org/10.1016/j.cois.2020.05.006 Eastwood G, Donnellycolt AK, Shepard JJ, Misencik MJ, Bedoukian R, Cole L, Armstrong PM, Andreadis TG (2020) Evaluation of novel trapping lures for monitoring exotic and native container-inhabiting Aedes spp. (Diptera: Culicidae) mosquitoes. J Med Entomol 57(2):534-541. Juarez, J. G., S. Garcia-Luna, L. F. Chaves, E. Carbajal, E. Valdez, C. Avila, W. Tang, E. Martin, R. Barrera, R. Hemme, J. P. Mutebi, N. Vuong, B. Roark, C. R. Maupin, I. E. Badillo-Vargas, G. L. Hamer. 2020. Dispersal of female and male Aedes aegypti from discarded container habitats using a stable isotope mark-capture study design in South Texas. Nature Scientific Reports. 10:6803. Kache PA, Eastwood G, Collins-Palmer K, Katz M, Falco RC, Bajwa WI, Armstrong PM, Andreadis TG, Diuk-Wasser MA (2020) Environmental determinants of Aedes albopictus abundance at a northern limit of its range in the United States. Am J Trop Med Hyg 102(2):436-447. Little EAH, Williams SC, Stafford KC 3rd, Linske MA, Molaei G. 2019. Evaluating the Effectiveness of An Integrated Tick Management Approach on Multiple Pathogen Infection in Ixodes scapularis Questing Nymphs and Larvae Parasitizing White-Footed Mice. Experimental and Applied Acarology. 80: 127–136.  Martin E, W. Tang, C. Briggs, H. Hopson, J. G. Juarez, S. Garcia Luna, M. Wise de Valdez, I. Badillo-Vargas, M. Borucki, M. Frank, G. L. Hamer. In press. Cell fusing agent virus (Flavivirus) infection in Aedes aegypti in Texas: seasonality, comparison by trap type, and individual viral loads. Archives of Virology. Molaei G, Little EAH. 2020. A Case of Morphological Anomalies in Amblyomma americanum(Acari: Ixodidae) Collected from Nature. Experimental and Applied Acarology 2020; 81: 279–285. Molaei G, Little EAH, Williams SC, Stafford KC. 2019. Bracing for the Worst–Range Expansion of the Lone Star Tick in the Northeastern United States. New England Journal of Medicine 2019; 381: 2189–2192. Molaei G, Mertins JW, Stafford III KC. 2020. Enduring Challenge of Invasive Ticks: Introduction of Amblyomma Oblongoguttatum(Acari: Ixodidae) into the United States on A Human Traveler Returning from Central America. Journal of Parasitology 2020; 106: 670–674. Olson M. F., S. Garcia-Luna, J. G. Juarez, E. Martin, L. C. Harrington, M. D. Eubanks, I. E. Badillo-Vargas, G. L. Hamer. In press. Sugar feeding patterns for Aedes aegypti and Culex quinquefasciatus (Diptera: Culicidae) mosquitoes in South Texas. Journal of Medical Entomology. Olson, M. F., M. L. Ndeffo-Mbah, J. G. Juarez, S. Garcia-Luna, E. Martin, M. K. Borucki, M. Frank, J. G. Estrada Franco, M. A. Rodriguez-Perez, N. A. Fernandez-Santos, G. J. Molina-Gamboa, S. D. Carmona Aguirre, B. L. Reyes-Berrones, L. J. Cortes-De la cruz, A. Garcia-Barrientos, R. E. Huidobro-Guevara, R. M. Brussolo-Ceballos, J. Ramirez, A. Salazar, L. F. Chaves, I. E. Badillo-Vargas, G. L. Hamer. 2020. High rate of non-human feeding by Aedes aegypti reduces Zika virus transmission in South Texas. Viruses. 12:453. Pokutnaya D, Molaei G, Weinberger DM, Vossbrinck CR, Diaz AJ. 2020. Prevalence of Infection and Co-infection and Presence of Rickettsial Endosymbionts in Ixodes scapularis (Acari: Ixodidae) in Connecticut, USA. Journal of Parasitology; 106: 30–37. Stafford III KC, Ridge GE, Molaei G, Zarb C, Bevilacqua P. 2020. Rabbit Bot Fly Furuncular, Tracheopulmonary, and Human Bot Fly Infestations in Connecticut (Oestridae: Cuterebrinae). Journal of Medical Entomology 58(1):114-120. doi: 10.1093/jme/tjaa181. Stafford III KC, Williams SC, van Oosterwijk JG, Linske MA, Zatechka S, Richer LM, Molaei G, Przybyszewski C, Wikel SK. Field Evaluation of a Novel Oral Reservoir‐Targeted Vaccine Against Borrelia burgdorferiUtilizing an Inactivated Whole‐Cell Bacterial Antigen Expression Vehicle. Experimental and Applied Acarology. 2020; 80: 257–268.

Impact Statements

Develop, test, implement, and encourage novel control (management) interventions that reduce transmission of human and animal diseases using environmental sound and scientifically-based approaches

Date of Annual Report: 12/13/2022

Report Information

Annual Meeting Dates: 10/20/2022 - 10/20/2022
Period the Report Covers: 09/20/2021 - 10/20/2022

Participants

Armstrong, Phil (Connecticut Agricultural Experiment Station); Cohnstaedt, Lee (USDA-ARS); Couret, Nelle (University of Rhode Island); Dobson, Stephen (University of Kentucky); Fonseca, Dina (Rutgers University); Gardner, Allison (University of Maine); Gloria-Soria, Andrea (Connecticut Agricultural Experiment Station); Hamer, Gabriel (Texas A&M University); Lee, Yoosook (University of Florida); Leisnham, Paul (University of Maryland); Machtinger, Erika (Pennsylvania State University); Meuti, Megan (The Ohio State University); Molaei, Goudarz (Connecticut Agricultural Experiment Station); Noden, Bruce (Oklahoma State University); Oliva Chavez, Adela (Texas A&M University); Paskewitz, Susan (University of Wisconsin); Piermarini, Peter (The Ohio State University); Reiskind, Michael (North Carolina State University); Short, Sarah (The Ohio State University); Silver, Kris (Kansas State University); Smith, Ryan (Iowa State University)

Brief Summary of Minutes

The annual meeting was held over Zoom on Thursday, October 20, 2022 from 12-5pm EST. Dr. Gardner organized and facilitated the meeting with input from prior chairs Dr. Leisnham and Dr. Reiskind. We decided to hold the meeting remotely with the approval of our NIFA advisor due to ongoing concerns about travel during the COVID-19 pandemic.

During the first half of the meeting, seven new members of the Multistate project since our last annual meeting gave presentations introducing their labs and their research themes that are aligned with the project. The presenters were as follows:

Dr. Adela Oliva Chavez, Department of Entomology, Texas A&M University

Dr. Megan Meuti, Department of Entomology, The Ohio State University

Dr. Yoosook Lee, Florida Medical Entomology Laboratory, University of Florida

Dr. Andrea Gloria-Soria, The Connecticut Agricultural Experiment Station

Dr. Erika Machtinger, Department of Entomology, Pennsylvania State University

Dr. Kristopher Silver, Department of Entomology, Kansas State University

Dr. Gillian Eastwood, Department of Entomology, Virginia Tech University

During the remainder of the meeting, we held two 45-minute breakout room sessions and a full group discussion of the future of the Multistate project. The first breakout room session focused on the three themes of the project to facilitate networking on these topics. Each of three rooms centered on 1) developing and strengthening effective surveillance and monitoring of disease vectors, 2) determining the ecology and geographic distribution of invasive and native disease vectors under changing environmental conditions, or 3) developing novel control and management interventions. The second breakout room session focused on discussion of what we want to gain from the Multistate project in the future. Topics discussed included sharing of data, specimens, and other resources for research projects (e.g., focused on population genetics of vector species or changing distributions of vector species) and potentially developing a review paper among the Multistate project team to cultivate the identity of this long-running Multistate project. We also discussed the merits of different locations for the next annual meeting. Participants regularly attend some combination of the Entomological Society of America annual meeting, the American Society of Tropical Medicine and Hygiene annual meeting, and the Society for Vector Ecology annual meeting. We discussed leading a hybrid meeting next year that would preserve the convenience and high attendance of virtual meetings while allowing for the social and networking benefits of an in-person meeting. Finally, we agreed on the plan to renew the Multistate project after the September 2024 end date.

Accomplishments

Dr. Phil Armstrong’s lab (Connecticut Agricultural Experiment Station) continued to conduct research on the vector biology, control, and ecology of US domestic arboviruses such as Powassan virus, West Nile virus and eastern equine encephalitis virus.  This includes recent work on the vector competence of human biting ticks for Powassan virus.  They show that three species- Ixodes scapularis, Amblyomma americanum, and Dermacentor variabilis were equally competent vectors of Powassan virus highlighting their potential role in the ecology and epidemiology of Powassan virus.  In another study, they partnered with mosquito control contractors to evaluate the impact of catch basin larvicide applications to reduce entomological metrics of West Nile virus risk.  Larvicide applications reduced pupal abundance and the prevalence of host-seeking adults but with no detectable impact on entomological risk metrics for WNV. Further research is needed to better determine the level of control needed to reduce WNV transmission risk.  They recently published a review paper on EEE virus to examine the major drivers of disease outbreaks in the northeastern United States. They are currently evaluating the phylogeography and movement of EEE virus to better understand the environmental and virological factors associated with the largest EEE outbreak in modern history during 2019.   Dr. Doug Brackney’s lab (Connecticut Agricultural Experiment Station) has been performing an active tick surveillance program in 40 paired sampling sites (5 in each of CT’s 8 counties). The sample counts presented here reflect total specimens collected during 2021 (data from 2022 is still being collected). Testing of 479 female and 500 nymphal I. scapularis ticks collected through 30 November 2021 found adult blacklegged ticks were infected with B. burgdorferi (52.9%), B. microti (16.9%), A. phagocytophilum (13.1%), B. miyamotoi (1.6%), and Powassan virus (0.82%). For nymphal blacklegged ticks, the results statewide were B. burgdorferi (21.6%), B. microti (8.4%), A. phagocytophilum (5.8%), B. miyamotoi (2.0%), and Powassan virus (0.0%).   Dr. Stephen Dobson’s lab (University of Kentucky) worked in collaboration with agents of the KY Department of Health to establish a networked system for field data collection, lab identification, molecular testing of pools, analysis, and data reporting. This system was successfully used for both tick and mosquito collections throughout the state. The software allows for linkage of samples to picture, audio and text notes, which are available to all participants, including field workers, lab personnel and data managers. The resulting data is automatically formatted for national databases, including VectorSurv, ArboNet, etc.   Dr. Gillian Eastwood’s lab (Virginia Tech) has conducted research on La Crosse virus ecology and pathogenicity (specifically, lineage III strains), phenology of container-breeding mosquitoes in Blacksburg, Virginia, and tick species diversity, ecology, and pathogen prevalence in southwest Virginia, with a focus on tick-borne viruses.   Dr. Vincenzo Ellis’s lab (University of Delaware) has been conducting population genetic analyses of Borrelia burgdorferi in ticks and small mammals and avian malaria pathogen identification in native birds in Delaware. They also have been developing molecular sequencing protocols for pathogen population genetic and phylogenetic studies.   Dr. Dina Fonseca’s lab (Rutgers University) has worked on multiple aims of the multistate project. Goal 1. First reports of soft tick Carios kelleyi (Cooley and Kohls), a parasite of bats, from New Jersey and Vermont. Although thought to be widespread in North America, the ecology of C. kelleyi is not well understood, despite reports of this species feeding on humans and its consequent potential as a disease vector.  The association of C. kelleyi with bat species that regularly roost in such as attics and barns, and recent isolations from this tick of pathogens capable of infecting humans, companion animals and livestock underscore the need for further studies of these bat ectoparasites. They also reported on larvae and nymphs of Ixodes scapularis recovered from big brown bats, Eptesicus fuscus (Palisot de Beauvois) (Chiroptera: Vespertilionidae), at four locations in rural New York State, USA. This is the first report of bats as hosts of I. scapularis. All Ixodes infested bats were injured and found on the ground, therefore, parasitism by I. scapularis was likely opportunistic. Nonetheless, the large number of pathogens known to be associated with bats and the frequency with which I. scapularis bites people suggest that this host-tick relationship is of at least potential epidemiological significance. Goal 2. From 2018-2020 in a collaboration with the NY City department of Health and the Monmouth Co. (NJ) Ticks and Tick-borne Diseases Program we characterized and reported a multi-year collection of the Gulf Coast tick, Amblyomma maculatum Koch (Acaridae: Ixodida: Ixodidae) in Staten Island, New York City (NYC) as well as their detection in Brooklyn, NYC, and in Atlantic and Cumberland counties in southern New Jersey, USA. Notably, they also reported a high prevalence in the ticks (~50%) of the human pathogen Rickettsia parkeri. In 2022 we detected an established population of this southern tick species in Salem, Co. NJ with similar infection levels with Rickettsia parkeri (54%). Goal 3. They examined the effect of forest thinning on ticks, a management tool used in the New Jersey Pinelands and elsewhere to improve forest health and resilience, mitigate wildfire risk, and manage for wildlife. They found that, on average, questing tick abundance was 92% lower in thinned as compared with unthinned sites. Of the four tick species collected in unthinned sites only one was collected in thinned sites. Prevalence of vector-borne pathogens was similar between treatments, but the significant and very large decline in tick abundance indicates a lower risk of tick encounters. Our results add to the growing evidence that landscape and forest management can reduce local tick abundance thereby reducing the risk for human tick-borne disease.    Dr. Allison Gardner’s lab (University of Maine) has progressed toward the aims of this project through two research avenues.  First, they continued ongoing work using field experiments to study overwinter survival of the blacklegged tick, a major disease vector species, in its northern range.  They also initiated a new study of blacklegged tick host-seeking behavior under different weather and climate conditions, and anticipate continuing this work over the next year.  Second, they continued an ongoing study of the spread of Zika virus in the western hemisphere via domestic and international tourism.  They extended concepts and tools developed under this project to analyze impacts of human movements on the spread of COVID as well.   Dr. Andrea Gloria-Soria’s lab (Connecticut Agricultural Experiment Station) continued to build a reference database of Aedes aegypti mosquitoes worldwide based on microsatellite markers and genome-wide SNPs. Such a database was used to track the origin of new introductions of this invasive species to Utah and Nebraska. They investigated the genetic diversity and population dynamics of Aedes albopictus in the Northeastern USA using microsatellites to better understand its invasion history and inform control strategies. They are also using mosquito collections from the Connecticut Mosquito Surveillance program to determine the role of each species in driving heartworm infections in the region, with a special focus on the invasive mosquito Aedes albopictus.   Dr. Gabriel Hamer’s lab (Texas A&M University) in several vector-borne disease systems continued to advance the understanding of vector ecology while evaluating different forms of vector control.  They have advanced understanding of South Texas populations of Aedes aegypti by understanding susceptibility to an insect growth regulator, they have measured their diel host-seeking activity, identified that Ae. aegypti is contributing to dog heartworm transmission, and evaluated two vector control tools for population suppression.  They have also studied vector ecology and pathogen infection for Culex spp. mosquitoes, ticks, and triatomines.   Dr. Laura Harrington’s lab (Cornell University) conducted research to address Multistate NE1943 Objectives 1,2 and 3.  For objective 1, they conducted entomological surveys coupled with knowledge attitudes and practices surveys to understand  tick borne disease risk and perceptions of ticks and risk in park visitors on Staten Island (Hassett et al. 2022) as well as how ticks are monitored across public health and vector control districts in the USA (Mader et al. 2021).  For objective 2, they investigated the host feeding biology of the invasive Asian tiger mosquito on Long Island (Fikrig et al. 2021) and assessed optimal surveillance methods for the invasive Asian Longhorned tick (Sherpa et al. 2021). For control and management of disease vectors (obj 3) they conducted a scoping review of Lyme disease provider-patient communication (Nesgros et al. 2021), evaluated susceptibilility of Lyme disease vector, Ixodes scapularis to permethrin in a long-term 4-poster treatment area on Shelter Island,  NY (Burtis et al. 2021a), evaluated the efficacy of methoprene for eastern equine encephalitis virus vector control in Massachusetts (Burtis et al. 2021b) and determined community-wide efficacy of WNV larval vector control practices in CT (McMillan et al. 2021).    Dr. Yoosook Lee’s lab (University of Florida) is aimed at (1) developing and strengthening effective surveillance and monitoring of disease vectors at local and regional scales, (2) determining the ecology and geographic distribution of invasive and native disease vectors, and (3) developing novel control and management interventions and test their impacts on the transmission of human and animal diseases. Lee obtained mosquito samples from multiple locations within Florida, Hawaii, and other Pacific Islands. EDDMaps database is constructed to curate mosquito collection data and some data curation has started. Whole genome sequencing data was collected from the natural populations to estimate population connectivity between locations and to inform various models aimed at modeling mosquito distribution and dispersal. This will be critical in assessing impacts of various mosquito control measures on the transmission of mosquito-borne diseases. Lee is also serving as the Program Director of the Mosquito BEACONS working group improving the invasive mosquito surveillance and control capacity in the Southern states including AL, FL, GA, MS, LA, NC, SC, TX, and PR.   Dr. Paul Leisnham’s lab (University of Maryland) tested the hypothesis that the contents of different urban containers alter the effects of competition from the invasive mosquito, Aedes albopictus, on resident Culex pipiens. They found that functional containers, and trash cans in particular, facilitated the persistence of C. pipiens despite the invasion of competitively superior A. albopictus, likely due to greater nutrient concentrations.   Dr. Shirley Luckhart’s lab (University of Idaho) studies molecular biology. Cutting-edge diagnostic assays for vector-borne disease samples (vectors, pathogens, hosts) are largely limited to research laboratories or a few commercial entities in the U.S., are expensive, and difficult to compare across laboratories due to use of platforms that vary significantly. Furthermore, assay capacity is typically limited to a few targets of interest (e.g., limited multi-plexing) or, if thousands of targets can be analyzed, this is most often from a single organism (e.g., RNA-seq), with the challenge that analyses of infecting pathogens are confounded by this approach. To address these issues, thel work with colleagues to develop an innovative next-generation sequencing platform for improved diagnostics for arthropods of medical importance. Specifically, they will leverage our expertise to adapt existing technology for a platform for mosquito- and tick-borne diseases that can be used to identify up to 100,000 targets in a single reaction with a single sample. The single reaction format can be used for arthropod, host, and pathogen analyses (or all three, in the case of bloodfed arthropods) to identify known pathogens, screen for novel pathogens, confirm the identity of the arthropod vector, discern population genetic variation in any of the organisms in the sample, identify drug resistance genotypes or genotypes associated with altered virulence in the associated pathogens, identify the hosts that were fed upon by the arthropods, identify geographic variation in host genetics, etc. There is no current resource like this in the U.S. Initially, they will develop a panel of biomarkers that can be used to screen for diverse pathogen groups for the major human-biting tick and mosquito species, along with known vector arthropods and hosts, with sufficient design flexibility to detect invasive pathogens, vector arthropods, and hosts. Methods and assay conditions will be optimized and tested collaboratively against known standards and previously characterized samples before being applied to field-collected specimens to assess limits of detection, sensitivity and specificity. From this, they will develop best-practice recommendations and support the adoption of standardized methods for diagnosis and testing for samples from across the U.S.   Dr. Megan Meuti’s lab (The Ohio State University) has been conducting mosquito surveillance throughout the fall, winter and spring to determine when mosquitoes enter and exit from their overwintering dormancy. They have also collected blood-fed mosquitoes throughout the spring, summer and fall to identify seasonal changes in host use that may contribute to West Nile virus transmission dynamics. The Meuti lab has also investigated how urban pollutants, like artificial light at night and higher temperatures associated with heat islands, affect mosquito dormancy in the lab and in the field. They have also evaluated how traditional and novel, trap-based approaches affect the abundance of mosquitoes and beneficial insects in urban areas. Finally, they are conducting basic research on the connection between the circadian clock and the hormonal pathways that regulate seasonal responses in mosquitoes, as well as differences in gene expression between biting (anautogenous) and non-biting (autogenous) mosquito variants to hopefully identify novel targets for control.   Dr. Goudarz Molaei’s lab (Connecticut Agricultural Experiment Station) focuses on the eco-epidemiology of mosquito- and tick-borne pathogens of human and veterinary health concern. He is elucidating the role of mosquito species in the transmission of West Nile and eastern equine encephalitis viruses and other arboviruses and the contribution of avian species as mosquito hosts to the maintenance and amplification of these viruses in the northeastern U.S. He is also directing the Connecticut Tick and Tick-borne Pathogen Surveillance Programs at the Agricultural Experiment Station, and conducting research on the ecology, biology, and vector-host-pathogen interactions of tick vectors of human pathogens of human and veterinary health concern.    Dr. Bruce Noden’s lab (Oklahoma State University) has worked to strengthen surveillance and monitoring of disease vectors at local and regional scales by focusing on: 1) what hosts mosquitoes that use eastern redcedar, an invasive tree in the Great Plains, are feeding on.  In addition to identifying a wide variety of hosts, they have found that newly blood fed mosquitoes will fly great distances to rest in ERC; and 2) the pathogens in fleas obtained from free-roaming domestic cats in central Oklahoma.  From a relatively small sample of fleas, they identified two Rickettsia species and three Bartonella species.  Additionally, an ear mite was positive for Bartonella. These results highlight the need for more focus on free-roaming domestic cats and their ectoparasites in the Great Plains region.   Dr. Adela Oliva Chavez’s lab (Texas A&M University) studied epigenetic mechanisms, which allow the regulation of gene expression without the modification of DNA coding sequences. Although several mechanisms of epigenetic regulation have been shown in ticks, their role in environmental and host adaptation have not been explored in depth. They currently are investigating two epigenetic mechanisms in the context of tick feeding and environmental adaptation: DNA methylation and miRNA expression. Their preliminary data indicates that methylation levels change in Ixodes scapularis populations from year to year, suggesting a potential mechanism for phenotypic plasticity. They have identified sites of differential methylation within their genome and pathway enrichment of genes differentially methylation. Lastly, they developed a new protocol for the isolation of miRNAs from tick salivary glands and extracellular vesicles, which reduces the number of ticks needed for experiments.   Dr. Reddy Palli’s lab (University of Kentucky) conducted surveillance for A. americanum in Kentucky through field collections and the establishment of a statewide tick submission program with the help of the Kentucky Department for Public Health and screened for Ehrlichia chaffeensis on a county-level throughout the state. They collected 5,726 A. americanum ticks in 77 counties and detected E. chaffeensis in 32 counties. The minimum infection rate was 1.8%. With the expansion of A. americanum and increasing cases of tick-borne diseases, future surveillance is needed to monitor this important tick vector over time.   Dr. Susan Paskewitz’s lab (University of Wisconsin) analyzed results of surveillance and control of blacklegged ticks and the Culex mosquitoes that transmit West Nile virus. They also completed studies on the role of microclimates in facilitating overwintering of Aedes albopictus. Finally, they focused on ecological studies of the small mammals associated with tickborne pathogens.   Dr. Dana Price’s lab (Rutgers University) focuses on functional genomic analyses of vector arthropods.  In 2021, they utilized Multistate resources for several such projects centered on ticks: Isolation and genome sequencing of two Rickettsial agents co-infecting  bat tick (Carios kelleyi) ticks discovered in New Jersey, and in a comparison of Ixodes scapularis (black-legged tick) populations and associated pathogens in two NJ counties. They also conducted similar research with mosquitoes that includes a tissue-tropic metaviromic assessment of single wild-caught mosquitoes using both Illumina and Oxford Nanopore sequencing technologies, and the generation of a draft genome of Anopheles bradleyi mosquito (a project that aims to sequence members of the Anopheles crucians complex). They remain very interested in emerging and under-studied insect vectors as well, and in 2022 generated a genome sequencing of the phlebotomine sand fly Lutzomyia vexator and associated novel Torix-group Rickettsial endosymbiont using Oxford Nanopore single-molecule sequencing.   Dr. Michael Reiskind’s lab (North Carolina State University) has conducted several studies as part of the multistate project.  They have on-going surveillance of invasive container mosquitoes (Aedes albopictus, Aedes aegypti, Aedes japonicas).  They also have on-going tick surveillance in North Carolina counties, in collaboration with NC Department of Health and Human Services, as well as providing entomological surveillance for epidemiological investigations of tick-borne disease.  They have also started three new research projects during this time frame.  The first focusing on the relative abundance of two important ticks in North Carolina: Amblyomma americanum and Dermacentor variabilis, comparing historical to contemporary data.  The second examines the role of larval habitat on response to parasitic infection in mosquitoes.  The final project, in collaboration with colleagues at East Carolina University and Western Carolina University, explores new technology (mid-infrared spectroscopy) to identify, age, and determine infection status of mosquitoes.     Dr. Kris Silver’s lab (Kansas State University) is exploring the use of RNA interference (RNAi), a new technology that can provide highly specific control of insect pests, as well as established insecticides like Bacillus thuringiensis israelensis as new avenues of control of Culicoides midges and Aedes mosquitoes. Additionally, they are also characterizing probing behaviors of Culex tarsalis and Culicoides midges using electropenetography to better understand how they feed and identify potential new targets for vector control.   Dr. Ryan Smith’s lab (Iowa State University) performed mosquito and tick surveillance in the state of Iowa to better understand West Nile virus transmission dynamics, the spread of invasive mosquito species, and to determine the current range of tick populations. When paired with the long-term data sets obtained from surveillance efforts in previous years, these data provide valuable insights into vector ecology and vector-borne disease transmission in the state of Iowa and the greater Midwest.   Dr. Alvaro Toledo’s lab (Rutgers University) studied natural commercial alternatives to synthetic repellents with similar or better properties than DEET. Tey evaluated the repellency of two extracts, CR3 and CR9, derived for newly developed catnip cultivars on two tick species, Ixodes scapularis and Haemaphysalis longicornis. Dose-response in vitro assays showed that CR3 and CR9 extracts have similar repellency properties to DEET. Few documented control strategies exist for the invasive tick, Haemaphysalis longicornis, despite its potential to reach extremely high numbers and vector human and animal pathogens. In 2020, they evaluated the effects of single applications of five granular and liquid acaricides on H. longicornis in a public park in northern New Jersey. Acaricides tested included pyrethroids (lambda-cyhalothrin, bifenthrin), a carbamate (carbaryl), and the insect growth regulators (IGRs) pyriproxyfen and novaluron. They also monitored the impact of each treatment on non-target soil and above-ground invertebrate species using pitfall and sticky traps, respectively.

Publications

Abernathy HA, BD Hollingsworth, DA Giandomenico, KA Moser, MH Reiskind, R Boyce. 2022.  Prevalence of Knock-Down Resistance F1534S Mutations in Aedes albopictus (Skuse)(Diptera: Culicidae) in North Carolina. Journal of Medical Entomology.   Adams, D. R.*, A. J. Golnar*, S. A. Hamer, M. A. Slotman, G. L. Hamer. 2021. Culex quinquefasciatus (Diptera: Culicidae) survivorship following the ingestion of bird blood infected with Haemoproteus sp. parasites. Parasitology Research. 120:2343–2350.   Adams, D. R., A. J. Golnar, J. I. Meyers, M. A. Slotman, G. L. Hamer. In press. Plasmodium relictum infection in Culex quinquefasciatus (Culicidae) decreases diel flight activity but increases peak dusk flight activity. Malaria Journal.   Adelman, J.S., Tokarz, R.E., Euken, A.E., Field, E.N., Russell, M.C., Smith, R.C. (2022) Relative influence of land use, mosquito abundance, and bird communities in defining West Nile virus infection rates in Culex mosquito populations. Insects 13 (9): 758.   Armstrong PM, Andreadis TG. (2022) Ecology and Epidemiology of Eastern Equine Encephalitis Virus in the Northeastern United States: An Historical Perspective. J Med Entomol 59(1):1-13.   Aryaprema VS, Qualls WA, Dobson KL, Dobson SL, Xue RD. The Effects of Boric Acid Sugar Bait on Wolbachia Trans-Infected Male Aedes albopictus (ZAP Males((R))) in Laboratory Conditions. Insects. 2021;13(1). Epub 20211221. doi: 10.3390/insects13010001. PubMed PMID: 35055844; PubMed Central PMCID: PMCPMC8777746.   Bajwa WI, Tsynman L, Egizi AM, Tokarz R, Maestas L, Fonseca DM 2022 The Gulf Coast tick, Amblyomma maculatum (Ixodida: Ixodidae) and spotted fever group Rickettsia in the highly urbanized northeastern US. Journal of Medical Entomology 59(4):1434-1442. https://doi.org/10.1093/jme/tjac053   Balasubramanian S, Curtis-Robles R, Chirra B, Auckland LD, Mai A, Bocanegra-Garcia V, Clark P, Clark W, Cottingham M, Fleurie G, Johnson CD, Metz RP, Wang S, Hathaway NJ, Bailey JA , Hamer GL, Hamer SA. 2022. Characterization of triatomine bloodmeal sources using direct Sanger sequencing and amplicon deep sequencing methods. Sci Rep 12:10234.   Beck A., Bjork J., Biggerstaff B., Eisen L., Eisen R., Foster E., Signs K., Tsao J., Kough E., Peterson M., Schiffman E., Muganda C., Osborn R., Wozniak R., Bron G., Phaneuf D., Smith D., Bartholomay L., Paskewitz S., Hinckley A.,  Knowledge, attitudes, and behaviors regarding tick-borne disease prevention in Lyme disease-endemic areas of the Upper Midwest, United States.  Ticks and Tickborne Disease. 13:101925. 2022. https://doi.org/10.1016/j.ttbdis.2022.101925   Brennan JR, Boychuck S, Washkwich AJ, John-Alder H, Fonseca DM 2022 Tick abundance and diversity is substantially lower in thinned vs. unthinned forests in the New Jersey Pinelands National Reserve, USA. Ticks and Tick-borne Diseases. Accepted with minor revisions (resubmitted)   Bron G., Fenelon H., and Paskewitz S.M. Assessing recognition of the vector of Lyme disease using resin-embedded specimens in a Lyme endemic area. Journal of Medical Entomology. 58:866-872. https://doi.org/10.1093/jme/tjaa234. 2021.   Bron G., Lee X. and Paskewitz S.M. Do-it-yourself tick control: granular gamma-cyhalothrin reduces Ixodes scapularis (Acari: Ixodidae) nymphs in residential backyards. Journal of Medical Entomology. 58:749-755.  https://doi.org/10.1093/jme/tjaa212.  2021.   Burtis JC, Bickerton MW, Indelicato N, Poggi JD, Crans SC and LC Harrington. 2022. Effectiveness of a Buffalo Turbine and A1 Mist Sprayer for the Areawide Deployment of Larvicide for Mosquito Control in an Urban Residential Setting. Journal of Medical Entomology.   Burtis JC, Poggi J, Duval, TB, Bidlack E, Shepard JJ, Matton P, Rossetti R, and LC Harrington. 2021. Evaluation of a methoprene aerial application for the control of Culiseta melanura (Diptera: Culicidae) in wetland breeding larval habitats. Journal of Medical Entomology. 58 (6), 2330-2337.   <a href="Burtis%20JC,%20Poggi%20J,%20Payne%20B,%20Campbell%20SR,%20and%20LC%20Harrington.%202021.%20%20The%20susceptibility%20of%20Ixodes%20scapularis%20collected%20from%20a%20long-term%204-poster%20treatment%20area%20on%20Shelter%20Island,%20Long%20Island%20NY.%20J%20Med%20Entomol.%20https:/doi.org/10.1093/jme/tjab054">Burtis JC, Poggi J, Payne B, Campbell SR, and LC Harrington. 2021.  The susceptibility of Ixodes scapularis collected from a long-term 4-poster treatment area on Shelter Island, Long Island NY. J Med Entomol. https://doi.org/10.1093/jme/tjab054</a>   Conte CE, JE Leahy, and AM Gardner. 2021. Active forest management reduces exposure risk to blacklegged ticks and tick-borne pathogens. EcoHealth 18: 157-168.   Crawford JE, Hopkins KC, Buchman A, Zha T, Howell P, Kakani E, et al. Reply to: Assessing the efficiency of Verily's automated process for production and release of male Wolbachia-infected mosquitoes. Nat Biotechnol. 2022. Epub 20220526. doi: 10.1038/s41587-022-01325-y. PubMed PMID: 35618925.   Cumbie AN, Whitlow AM, Eastwood G (2021) First Evidence of Powassan Virus (Flaviviridae) in Ixodes scapularis in Appalachian Virginia, USA. AJTMH 103(3): 905-908. doi.org/10.4269/ajtmh.21-0825   Dacso, M. M., D. A. Bente, S. C. Weaver, G. P. Kobinger, P. C. Melby, S. L.F. McLellan, P. H. Keiser, S. A. Hamer, G. L. Hamer, G. W. Parker, D. I. Douphrate, A. Rodriguez, M. L. Goodman, G. C. Gray. 2022. Texas professionals are employing a one health approach to protect the United States against biosecurity threats. One Health. 15:100431.   Davila, E., N. A. Fernandez-Santos, J. G. Estrada-Franco, L. Wei, J. A. Agular-Duran, M. J. Lopez-Lopez, R. Solis- Hernandez, R. Garcia-Miranda, D. D. Valazquez-Ramirez, J. Torres-Romero, S. Arellano Chavez, R. Cruz-Cadena, R. Navarro-Lopez, A. A. Perez de Leon, C. Guichard-Romero, E. Martin, W. Tang, M. Frank, M. Borucki, M. J. Turell, A. Pauvolid-Correa, M. A. Rodriguez-Perez, H. Ochoa-Diaz-Lopez, S. A. Hamer, G. L. Hamer. 2022. Utility of domestic dogs as effective sentinels for WNV transmission, but not Aedes-borne flavivirus transmission, in Mexico.  Emerging Infectious Diseases. 28: 1071–1074.   De Nadai BL, AG Maletzke, JJ Corbi, G Batista, MH Reiskind. 2021.  The impact of body size on Aedes [Stegomyia] aegypti wingbeat frequency: implications for mosquito identification.  Medical and Veterinary Entomology 35 (4), 617-624   Dickinson, K. L., N. Banacos, E. Carbajal, N. Dacko, C. Fredregill, S. Hinojosa, J. G. Juarez*, C. Weldon, G. L. Hamer. 2022. Public willingness to pay and social acceptability for mosquito control in Texas. Emerging Infectious Diseases. 28: 425-428.   Dobson KL, Mains JW, Dobson SL. Sterile Insect Technique (SIT) for Individual Property Owners: An Overview of the MosquitoMate Experience. Wing Beats. 2022;33(Spring).   Dobson SL. When More is Less: Mosquito Population Suppression Using Sterile, Incompatible and Genetically Modified Male Mosquitoes. J Med Entomol. 2021;58(5):1980-6. Epub 2021/03/12. doi: 10.1093/jme/tjab025. PubMed PMID: 33704487.   Dong, B., L. Khan, M. Smith, J. Trevino, M. Zhao, G. L. Hamer, A. Lemus, A. A. Molina, J. Lubinda, U. DT. Nguyen, U. Haque. Spatio-temporal dynamics of chikungunya, dengue, and Zika viruses in Mexico. In press. Communications Medicine.   Elias SP, AM Gardner, KA Maasch, SD Birkel, NT Anderson, PW Rand, CB Lubelczyk, and RP Smith. 2021. A generalized additive model correlating blacklegged ticks with white-tailed deer density, temperature, and humidity in Maine, USA, 1990-2013. Journal of Medical Entomology 58: 125-138.   Ellis VA, V Kalbskopf, A Ciloglu, M Duc, X Huang, A Inci, S Bensch, O Hellgren, V Palinauskas. 2022. Genomic sequence capture of Plasmodium relictum in experimentally infected birds. Parasites and Vectors 15: 1-12.   Field, E.N., Shepard, J. J., Clifton, M. E., Price, K. J., Witmier, B. J., Johnson, K., Boze, B., Abadam, C., Ebel, G. D., Armstrong, P.M., Barker, C. M., Smith, R. C. (2022) Semi-field and surveillance data define the natural diapause timeline for Culex pipiens across the United States. bioRxiv 2022.05.19.492729.   Figurskey AC, B Hollingsworth, MS Doyle, MH Reiskind. 2022. Effectiveness of autocidal gravid trapping and chemical control in altering abundance and age structure of Aedes albopictus.  Pest Management Science   Fikrig K, Peck S, Deckerman P, Dang S, St Fleur K, Goldsmith H, Qu S, Rosenthal H and LC Harrington. 2021. The effects of host availability and fitness on Aedes albopictus blood feeding patterns in New York. American Journal of Tropical Medicine and Hygiene. 106(1), 320-331.   Foster E., Burtis J., Tsao J., Bjork J, Liu G., Nietzel D.F., Schiffman E., Lee X., Paskewitz S., Caporale D., Eisen R.J. Inter-annual variation in prevalence of Borrelia burgdorferi sensu stricto and Anaplasma phagocytophilum in host-seeking Ixodes scapularis (Acari: Ixodidae) at long-term surveillance sites in the upper midwestern United States: Implications for public health practice. Ticks and Tick-borne Disease. 13: 101886. 2022.  https://doi.org/10.1016/j.ttbdis.2021.101886   Frederick, J.C., Sharma, P., Thompson, A., Dharmarajan, G., Ronai, I., Pesapane, R., Smith, R.C., Sundstrom, K., Tsao, J.I., Tuten, H., Yabsley, M.J., Glenn, T.C. (2022) Phylogeography of the blacklegged tick Ixodes scapularis identifies candidate loci for differences in vectorial capacity. Authorea. August 23, 2022.   Gallagher, M. R., J. Kreye, E. T. Machtinger, N. Schmidt, A. Everland, N. S. Skowronski. 2022. Can restoration of fire-dependent ecosystems reduce ticks and tick-borne disease prevalence in the Eastern US? Ecological Applications. 32: e2637 https://doi.org/10.1002/eap.2637   Gloria-Soria, A., 2022. 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