Assessing the value and knowledge gains from an online tick identification and tick-borne disease management course for the Southeastern United States.

BACKGROUND: Tick-borne diseases are a growing public health threat in the United States. Despite the prevalence and rising burden of tick-borne diseases, there are major gaps in baseline knowledge and surveillance efforts for tick vectors, even among vector control districts and public health agencies. To address this issue, an online tick training course (OTTC) was developed through the Southeastern Center of Excellence in Vector-Borne Diseases (SECOEVBD) to provide a comprehensive knowledge base on ticks, tick-borne diseases, and their management. METHODS: The OTTC consisted of training modules covering topics including tick biology, tick identification, tick-borne diseases, and public health, personal tick safety, and tick surveillance. The course was largely promoted to vector control specialists and public health employees throughout the Southeastern US. We collected assessment and survey data on participants to gauge learning outcomes, perceptions of the utility of knowledge gained, and barriers and facilitators to applying the knowledge in the field. RESULTS: The OTTC was successful in increasing participants' baseline knowledge across all course subject areas, with the average score on assessment increasing from 62.6% (pre-course) to 86.7% (post-course). More than half of participants (63.6%) indicated that they would definitely use information from the course in their work. Barriers to using information identified in the delayed assessment included lack of opportunities to apply skills (18.5%) and the need for additional specialized training beyond what the OTTC currently offers (18.5%), while the main facilitator (70.4%) for applying knowledge was having opportunities at work, such as an existing tick surveillance program. CONCLUSIONS: Overall, this OTTC demonstrated capacity to improve knowledge in a necessary and underserved public health field, and more than half of participants use or plan to use the information in their work. The geographic reach of this online resource was much larger than simply for the Southeastern region for which it was designed, suggesting a much broader need for this resource. Understanding the utility and penetrance of training programs such as these is important for refining materials and assessing optimal targets for training.

Conserving spawning stocks through harvest slot limits and no-take protected areas.

The key to the conservation of harvested species is the maintenance of reproductive success. Yet for many marine species large, old, individuals are targeted despite their disproportionate contribution to reproduction. We hypothesized that a combination of no-take marine protected areas (MPAs) and harvest slot limits (maximum and minimum size limits) would result in the conservation of large spawning individuals under heavy harvest. We tested this approach under different harvest intensities with a 2-sex, stage-structured metapopulation model for the Caribbean spiny lobster (Panulirus argus). P. argus is intensively harvested in the Caribbean, and in many localities large, mature individuals no longer exist. No-take MPAs and harvest slot limits combined, rebuilt and maintained large mature individuals even under high harvest pressure. The most conservative model (a 30% MPA and harvest slot limit of 75-105 mm) increased spawner abundance by 5.53E12 compared with the fishing status quo at the end of 30 years. Spawning stock abundance also increased by 2.76-9.56E12 individuals at a high harvest intensity over 30 years with MPAs alone. Our results demonstrate the potential of MPAs and harvest slot limits for the conservation of large breeding individuals in some marine and freshwater environments. Decisions on which management strategy best suits a fishery, however, requires balancing what is ecologically desirable with what is economically and socially feasible.

Conservación de Stocks Reproductivos por medio de Límites de Espacios de Producción y Áreas Protegidas de Cero Captura Resumen La clave para la conservación de las especies en criaderos es la manutención del éxito reproductivo. Aún así, para muchas especies marinas los individuos grandes y viejos son seleccionados a pesar de su contribución desproporcionada para la reproducción. Nuestra hipótesis supone que una combinación de áreas marinas protegidas (AMPs) de cero captura y los límites de espacio de producción (límites máximos y mínimos de tamaño) resultaría en la conservación de individuos reproductivos grandes durante una producción intensiva. Probamos esta estrategia bajo diferentes intensidades de crianza con un modelo metapoblacional de dos sexos y estructurado por etapas aplicado a la langosta espinosa del Caribe (Panulirus argus). P. argus es producida intensivamente en el Caribe y en muchas de las localidades, los individuos grandes y maduros ya no existen. La combinación de las AMPs de cero captura y los límites de espacio de producción regeneró y mantuvo individuos grandes y maduros incluso bajo una presión alta de producción. El modelo más conservador (un 30% de MPA y un límite de espacio de producción de 75-105 mm) incrementó la abundancia de individuos reproductivos por 5.53E12 comparado con el status quo de la pesca después de treinta años. La abundancia del stock reproductivo también incrementó por 2.76-9.56E12 individuos en una intensidad alta de producción durante 30 años sólo con las AMPs. Nuestros resultados demuestran el potencial de las AMPs y los límites de espacio de producción para la conservación de individuos reproductivos grandes en algunos ambientes marinos y de agua dulce. Sin embargo, las decisiones sobre cuál es la mejor estrategia de manejo para una pesquería requieren del equilibrio entre lo que es ecológicamente deseable y lo que es económica y socialmente factible.

A Simple, Inexpensive Method for Mark-Recapture of Ixodid Ticks.

Mark-recapture techniques have been widely used and specialized to study organisms throughout the field of biology. To mark-recapture ticks (Ixodida), we have created a simple method to mark ticks using nail polish applied with an insect pin secured in a pencil that allows for a variety of questions to be answered. For measuring tick control efficacy, estimating population estimates, or measuring movement of ticks, this inexpensive mark-recapture method has been easily applied in the field and in the lab to provide useful data to answer a variety of questions about ticks.

A stochastic epidemic model for the dynamics of two pathogens in a single tick population.

Understanding tick-transmitted pathogens in tick infested areas is crucial for the development of preventive and control measures in response to the increasing cases of tick-borne diseases. A stochastic model for the dynamics of two pathogens, Rickettsia parkeri and Rickettsia amblyommii, in a single tick, Amblyomma americanum, is developed and analysed. The model, a continuous-time Markov chain, is based on a deterministic tick-borne disease model. The extinction threshold for the stochastic model is computed using the multitype branching process and conditions for pathogen extinction or persistence are presented. The probability of pathogen extinction is computed using numerical simulations and is shown to be a good estimate of the probability of extinction calculated from the branching process. A sensitivity analysis is undertaken to illustrate the relationship between co-feeding and transovarial transmission rates and the probability of pathogen extinction. Expected epidemic duration is estimated using sample paths and we show that R. amblyommii is likely to persist slightly longer than R. parkeri. Further, we estimate the duration of possible coexistence of the two pathogens.

Rickettsia parkeri infections diagnosed by eschar biopsy, Virginia, USA.

BACKGROUND: Infection with Rickettsia parkeri is an emerging tick-borne illness, often accompanied by fever and an eschar at the site of tick attachment. We present three cases of R. parkeri in Virginia residents. CASE PRESENTATIONS: Case 1 presented initially afebrile, failed to seroconvert to rickettsial antigens, and was diagnosed by DNA testing of the eschar. Case 2 presented febrile with eschar, no serologies were performed, and was diagnosed by DNA testing of the eschar. Case 3 presented febrile with eschar, serologies were negative for rickettsial antigens, and was diagnosed by DNA testing of the eschar. CONCLUSION: DNA testing of eschars represents an under-utilized diagnostic test and may aid in cases where the diagnosis is not made clinically.

A Stochastic Tick-Borne Disease Model: Exploring the Probability of Pathogen Persistence.

We formulate and analyse a stochastic epidemic model for the transmission dynamics of a tick-borne disease in a single population using a continuous-time Markov chain approach. The stochastic model is based on an existing deterministic metapopulation tick-borne disease model. We compare the disease dynamics of the deterministic and stochastic models in order to determine the effect of randomness in tick-borne disease dynamics. The probability of disease extinction and that of a major outbreak are computed and approximated using the multitype Galton-Watson branching process and numerical simulations, respectively. Analytical and numerical results show some significant differences in model predictions between the stochastic and deterministic models. In particular, we find that a disease outbreak is more likely if the disease is introduced by infected deer as opposed to infected ticks. These insights demonstrate the importance of host movement in the expansion of tick-borne diseases into new geographic areas.

Invasion of two tick-borne diseases across New England: harnessing human surveillance data to capture underlying ecological invasion processes.

Modelling the spatial spread of vector-borne zoonotic pathogens maintained in enzootic transmission cycles remains a major challenge. The best available spatio-temporal data on pathogen spread often take the form of human disease surveillance data. By applying a classic ecological approach-occupancy modelling-to an epidemiological question of disease spread, we used surveillance data to examine the latent ecological invasion of tick-borne pathogens. Over the last half-century, previously undescribed tick-borne pathogens including the agents of Lyme disease and human babesiosis have rapidly spread across the northeast United States. Despite their epidemiological importance, the mechanisms of tick-borne pathogen invasion and drivers underlying the distinct invasion trajectories of the co-vectored pathogens remain unresolved. Our approach allowed us to estimate the unobserved ecological processes underlying pathogen spread while accounting for imperfect detection of human cases. Our model predicts that tick-borne diseases spread in a diffusion-like manner with occasional long-distance dispersal and that babesiosis spread exhibits strong dependence on Lyme disease.

New Records of Ixodes affinis (Acari: Ixodidae) Parasitizing Avian Hosts in Southeastern Virginia.

Ixodes affinis Neumann (Acari: Ixodidae) is a hard-bodied tick species distributed throughout much of the southeastern United States. Although I. affinis does not parasitize humans, it is a competent vector of Borrelia burgdorferi sensu stricto, the causative-agent of Lyme disease, and thus contributes to the enzootic maintenance of this pathogen. This study presents evidence of I. affinis parasitizing five new host passerine species. During 2012-2014, 1,888 birds were captured and examined for ticks, and 18 immature I. affinis were collected from 12 birds-six Carolina Wrens (Thyrothorus ludovicianus); two Brown Thrashers (Toxostoma rufum); and one American Robin (Turdus migratorius), Eastern Towhee (Pipilo erythrophthalmus), Northern Cardinal (Cardinalis cardinalis), and White-throated Sparrow (Zonotrichia albicollis). Of 15 larvae and 3 nymphs collected, one nymph tested positive for B. burgdorferi DNA. I. affinis was found co-feeding on birds with immature Amblyomma americanum (L.), Ixodes brunneus Koch, Ixodes dentatus Marx, Ixodes scapularis Say, and Haemaphysalis leporispalustris Packard. The results of this research provide a better understanding of I. affinis hosts and identify avian taxa that may play a role in the maintenance and dispersal of this tick species.

Rickettsia parkeri Transmission to Amblyomma americanum by Cofeeding with Amblyomma maculatum (Acari: Ixodidae) and Potential for Spillover.

Amblyomma americanum (L.) is a human-biting ixodid tick distributed throughout much of the southeastern United States. Rickettsia parkeri is a member of the spotted fever group rickettsiae and causes a febrile illness in humans commonly referred to as "Tidewater spotted fever" or "R. parkeri rickettsiosis." Although the Gulf Coast tick, Amblyomma maculatum Koch, is the primary vector of R. parkeri, a small proportion of A. americanum have also been shown to harbor R. parkeri. The purpose of this investigation was to determine whether R. parkeri is spilling over into A. americanum in eastern Virginia and also to determine through laboratory experiments, whether A. americanum can acquire R. parkeri by cofeeding alongside infected ticks. Of 317 wild-caught, flat adult A. americanum tested from 29 counties and independent cities in coastal Virginia, a single female A. americanum was positive for R. parkeri, suggesting that R. parkeri is spilling over into this species, but at very low rates (<1.0%). Laboratory studies using guinea pigs indicated that nymphal A. americanum were able to acquire R. parkeri while feeding alongside infected A. maculatum and then transstadially maintain the infection. Nymphal A. americanum infected with Rickettsia amblyommii, however, were less likely to acquire R. parkeri, suggesting that infection with R. amblyommii may prevent R. parkeri from establishing infection in A. americanum.

Estimating the reproductive numbers for the 2008-2009 cholera outbreaks in Zimbabwe.

Cholera remains an important global cause of morbidity and mortality, capable of causing periodic epidemic disease. Beginning in August 2008, a major cholera epidemic occurred in Zimbabwe, with 98,585 reported cases and 4,287 deaths. The dynamics of such outbreaks, particularly in nonestuarine regions, are not well understood. We explored the utility of mathematical models in understanding transmission dynamics of cholera and in assessing the magnitude of interventions necessary to control epidemic disease. Weekly data on reported cholera cases were obtained from the Zimbabwe Ministry of Health and Child Welfare (MoHCW) for the period from November 13, 2008 to July 31, 2009. A mathematical model was formulated and fitted to cumulative cholera cases to estimate the basic reproductive numbers R(0) and the partial reproductive numbers from all 10 provinces for the 2008-2009 Zimbabwe cholera epidemic. Estimated basic reproductive numbers were highly heterogeneous, ranging from a low value of just above unity to 2.72. Partial reproductive numbers were also highly heterogeneous, suggesting that the transmission routes varied by province; human-to-human transmission accounted for 41-95% of all transmission. Our models suggest that the underlying patterns of cholera transmission varied widely from province to province, with a corresponding variation in the amenability of outbreaks in different provinces to control measures such as immunization. These data underscore the heterogeneity of cholera transmission dynamics, potentially linked to differences in environment, socio-economic conditions, and cultural practices. The lack of traditional estuarine reservoirs combined with these estimates of R(0) suggest that mass vaccination against cholera deployed strategically in Zimbabwe and surrounding regions could prevent future cholera epidemics and eventually eliminate cholera from the region.

Release the hens: a study on the complexities of guinea fowl as tick control.

Established tick control strategies often involve methods that can be damaging to existing environmental conditions or natural host ecology. To find more environmentally friendly methods, biological controls, like predators of ticks, have been suggested. There are natural predators of ticks, but most are generalists and not expected to control tick populations. Helmeted guinea fowl (Numida meleagris (L.) (Galliformes: Numididae)) have been suggested to be biological controls of ticks, and therefore, tick-borne pathogens, but their potential role as hosts for ticks complicates the relationship. A study was conducted to assess whether guinea fowl reduces the abundance of lone star ticks, Amblyomma americanum (L.) (Acari: Ixodidae), or whether they are hosts of ticks. Using mark-recapture techniques, painted lone star ticks were placed into 3 different treatments: penned, excluded, and free range. The recapture rates of painted ticks were compared. There was a significant difference between excluded and free-range treatments, but not between excluded and penned or between free range and penned. To investigate the role of guinea fowl as hosts of ticks, coop floors were examined for engorged ticks. Engorged lone star nymphs that had fed on guinea fowl were found. Lastly, ticks collected were tested to identify the potential reduction in risk of tick-borne pathogens. This study found no evidence that guinea fowl are an effective biological control of lone star ticks or tick-borne pathogens, but they are hosts of lone star nymphs. Future studies are needed to assess the complex ecology of a biological control of ticks that is also a host.

Survival of adult Amblyomma americanum, Amblyomma maculatum, and Dermacentor variabilis (Acari: Ixodidae) in four habitats in southeastern Virginia.

Adult, unfed Amblyomma americanum (Linnaeus), Amblyomma maculatum Koch, and Dermacentor variabilis (Say) ticks were housed inside environmental containers in situ from May through August 2015. The environmental containers were placed in 4 habitats in southeastern Virginia, United States. Two of the habitats were located in low ground and were prone to flooding; the other two consisted of a drier landscape located in a more upland habitat. A Cox Regression survival analysis indicated there was a significant difference in survival among species across all field sites. There was a 50.5-times higher risk of mortality for A. maculatum compared to A. americanum, a 4.3-times higher risk of mortality for A. maculatum compared to D. variabilis, and an 11.9-times higher risk of mortality for D. variabilis compared to A. americanum. There was also significantly higher mortality in field sites prone to flooding than in drier, upland field sites. We concluded that A. americanum was not negatively affected by increased flooding or the variable environmental conditions in southeastern Virginia. Dermacentor variabilis also was successful at remaining in the environment off-host, although increased flooding reduced survival over time. Amblyomma maculatum was more susceptible to mortality during long periods of time off-host in both environments, with high mortality rates in both dry upland and low-ground, flood-prone habitats.

Newer Surveillance Data Extends our Understanding of the Niche of Rickettsia montanensis (Rickettsiales: Rickettsiaceae) Infection of the American Dog Tick (Acari: Ixodidae) in the United States.

Background: Understanding the geographic distribution of Rickettsia montanensis infections in Dermacentor variabilis is important for tick-borne disease management in the United States, as both a tick-borne agent of interest and a potential confounder in surveillance of other rickettsial diseases. Two previous studies modeled niche suitability for D. variabilis with and without R. montanensis , from 2002-2012, indicating that the D. variabilis niche overestimates the infected niche. This study updates these, adding data since 2012. Methods: Newer surveillance and testing data were used to update Species Distribution Models (SDMs) of D. variabilis , and R. montanensis infected D. variabilis , in the United States. Using random forest (RF) models, found to perform best in previous work, we updated the SDMs and compared them with prior results. Warren's I niche overlap metric was used to compare between predicted suitability for all ticks and 'pathogen positive niche' models across datasets. Results: Warren's I indicated <2% change in predicted niche, and there was no change in order of importance of environmental predictors, for D. variabilis or R. montanensis positive niche. The updated D. variabilis niche model overpredicted suitability compared to the updated R. montanensis positive niche in key peripheral parts of the range, but slightly underpredicted through the northern and midwestern parts of the range. This reinforces previous findings of a more constrained pathogen-positive niche than predicted by D. variabilis records alone. Conclusions: The consistency of predicted niche suitability for D. variabilis in the United States, with the addition of nearly a decade of new data, corroborates this is a species with generalist habitat requirements. Yet a slight shift in updated niche distribution, even of low suitability, included more southern areas, pointing to a need for continued and extended monitoring and surveillance. This further underscores the importance of revisiting vector and vector-borne disease distribution maps.

Newer Surveillance Data Extends our Understanding of the Niche of Rickettsia montanensis (Rickettsiales: Rickettsiaceae) Infection of the American Dog Tick (Acari: Ixodidae) in the United States.

Background: Understanding the geographic distribution of Rickettsia montanensis infections in Dermacentor variabilis is important for tick-borne disease management in the United States, as both a tick-borne agent of interest and a potential confounder in surveillance of other rickettsial diseases. Two previous studies modeled niche suitability for D. variabilis with and without R. montanensis, from 2002 to 2012, indicating that the D. variabilis niche overestimates the infected niche. This study updates these, adding data since 2012. Methods: Newer surveillance and testing data were used to update Species Distribution Models (SDMs) of D. variabilis, and R. montanensis-infected D. variabilis, in the United States. Using random forest models, found to perform best in previous work, we updated the SDMs and compared them with prior results. Warren's I niche overlap metric was used to compare between predicted suitability for all ticks and "R. montanensis-positive niche" models across datasets. Results: Warren's I indicated <2% change in predicted niche, and there was no change in order of importance of environmental predictors, for D. variabilis or R. montanensis-positive niche. The updated D. variabilis niche model overpredicted suitability compared with the updated R. montanensis-positive niche in key peripheral parts of the range, but slightly underpredicted through the northern and midwestern parts of the range. This reinforces previous findings of a more constrained R. montanensis-positive niche than predicted by D. variabilis records alone. Conclusions: The consistency of predicted niche suitability for D. variabilis in the United States, with the addition of nearly a decade of new data, corroborates this is a species with generalist habitat requirements. Yet a slight shift in updated niche distribution, even of low suitability, included more southern areas, pointing to a need for continued and extended monitoring and surveillance. This further underscores the importance of revisiting vector and vector-borne disease distribution maps.

Estimating the distribution of Oryzomys palustris, a potential key host in expanding rickettsial tick-borne disease risk.

Increasingly, geographic approaches to assessing the risk of tick-borne diseases are being used to inform public health decision-making and surveillance efforts. The distributions of key tick species of medical importance are often modeled as a function of environmental factors, using niche modeling approaches to capture habitat suitability. However, this is often disconnected from the potential distribution of key host species, which may play an important role in the actual transmission cycle and risk potential in expanding tick-borne disease risk. Using species distribution modeling, we explore the potential geographic range of Oryzomys palustris, the marsh rice rat, which has been implicated as a potential reservoir host of Rickettsia parkeri, a pathogen transmitted by the Gulf Coast tick (Amblyomma maculatum) in the southeastern United States. Due to recent taxonomic reclassification of O. palustris subspecies, we reclassified geolocated collections records into the newer clade definitions. We modeled the distribution of the two updated clades in the region, establishing for the first time, range maps and distributions of these two clades. The predicted distribution of both clades indicates a largely Gulf and southeastern coastal distribution. Estimated suitable habitat for O. palustris extends into the southern portion of the Mid-Atlantic region, with a discontinuous, limited area of suitability in coastal California. Broader distribution predictions suggest potential incursions along the Mississippi River. We found considerable overlap of predicted O. palustris ranges with the distribution of A. maculatum, indicating the potential need for extended surveillance efforts in those overlapping areas and attention to the role of hosts in transmission cycles.

Tick Species Composition, Collection Rates, and Phenology Provide Insights into Tick-Borne Disease Ecology in Virginia.

To better understand tick ecology in Virginia and the increasing Lyme disease incidence in western Virginia, a comparative phenological study was conducted in which monthly collections were performed at twelve sampling locations in southwestern Virginia (high Lyme disease incidence) and 18 equivalent sampling locations in southeastern Virginia (low Lyme disease incidence) for one year. In western Virginia, we also explored the effect of elevation on collection rates of Ixodes scapularis Say (Acari: Ixodidae) and Amblyomma americanum (L.) (Acari: Ixodidae). In total, 35,438 ticks were collected (33,106 A. americanum; 2,052 I. scapularis; 134 Ixodes affinis Neumann [Acari: Ixodidae]; 84 Dermacentor variabilis [Say] [Acari: Ixodidae]; 49 Dermacentor albipictus [Packard] [Acari: Ixodidae]; 10 Haemaphysalis leporispalustris [Packard] [Acari: Ixodidae]; 2 Ixodes brunneus Koch [Acari: Ixodidae]; 1 Haemaphysalis longicornis Neumann [Acari: Ixodidae]). Within southwestern Virginia, Ixodes scapularis collection rates were not influenced by elevation, unlike A. americanum which were collected more frequently at lower elevations (e.g., below 500 m). Notably, I. scapularis larvae and nymphs were commonly collected in southwestern Virginia (indicating that they were questing on or above the leaf litter) but not in southeastern Virginia. Questing on or above the leaf litter is primarily associated with northern populations of I. scapularis. These findings may support the hypothesis that I. scapularis from the northeastern United States are migrating into western Virginia and contributing to the higher incidence of Lyme disease in this region. This comparative phenological study underscores the value of these types of studies and the need for additional research to further understand the rapidly changing tick-borne disease dynamics in Virginia.

Estimating the impact of human papillomavirus (HPV) vaccination on HPV prevalence and cervical cancer incidence in Mali.

Human papillomavirus vaccines have potential to reduce cervical cancer incidence and mortality; however, cultural and economic barriers may hinder success in developing countries. We assessed impact of a single vaccine campaign in Mali with use of mathematical modeling. Our model shows that decreases in the prevalence of Human papillomavirus infection are proportional to achieved vaccination coverage.

Rickettsia parkeri in gulf coast ticks, southeastern Virginia, USA.

We report evidence that Amblyomma maculatum tick populations are well established in southeastern Virginia. We found that 43.1% of the adult Gulf Coast ticks collected in the summer of 2010 carried Rickettsia parkeri, suggesting that persons living in or visiting southeastern Virginia are at risk for infection with this pathogen.

Spatio-temporal modelling of tick life-stage count data with spatially varying coefficients.

There is a vast amount of geo-referenced data in many fields of study including ecological studies. Geo-referencing is usually by point referencing; that is, latitudes and longitudes or by areal referencing, which includes districts, counties, states, provinces and other administrative units. The availability of large geo-referenced datasets for modelling has necessitated the development and application of spatial statistical methods. However, spatial varying coefficients models exploring the abundance of tick counts remain limited. In this study we used data that was collected and prepared by researchers in the Department of Biological Sciences from the Old Dominion University, Virginia, USA. We modelled tick life-stage counts and abundance variability from 12 sampling locations, with 5 different habitats (numbered 1-5), three habitat types; namely: woods, edges and grass; collected monthly from May 2009 through December 2018. Spatio-temporal Poisson and spatio-temporal negative binomial (NB) count data models were fitted to the data and compared using the deviance information criteria (DIC). The NB model outperformed the Poisson models with all its DIC values being smaller than those of the Poisson model. Results showed that the covariates varied spatially across counties. There was a decreasing time (in years) effect over the study period. However, even though the time effect was decreasing over the study period, space-time interaction effects were seen to be increasing over time in York County.

Focus Stacking Images of Morphological Character States for Differentiating the Adults of Ixodes affinis and Ixodes scapularis (Acari: Ixodidae) in Areas of Sympatry.

Adult females and males of Ixodes affinis and Ixodes scapularis are illustrated by focus stacking image photography, and morphological character states are described that reliably differentiate the two species. In conjunction with other environmental cues, such as the questing phenology of adults, these characteristics will enable the rapid identification of adults of either sex along the southern Coastal Plain of the United States, where these species are sympatric.