Questing Behavior Research Articles

Electric transportation and electroreception in hummingbird flower mites

Electric fields in terrestrial environments are used by caterpillars to detect their predators, as foraging cues by pollinators, and facilitate ballooning by spiders. This study shows that electric fields facilitate transportation and detection of hummingbirds in a guild of tropical phoretic mites. Hummingbird flower mites feed on nectar and pollen and complete their life cycle inside flowers. Mites colonize new flowers by hitching rides on hummingbird beaks. Flower mites emerge from hummingbird nostrils and disembark when the beak touches a flower. We tested whether flower mites are attracted to unmodulated electrostatic, or to modulated electric fields with amplitudes and frequencies in the range of those previously reported for hummingbirds. In a laboratory setup, mites were only attracted to modulated electric fields. In a choice experiment between positive or negative polarities, mites almost instantaneously chose positive charges, but only when the field was modulated. Mites display questing behavior, moving their front legs toward an electrostatic source. In experiments where we removed one or both front leg tarsi, we show that modulated fields are detected by sensory structures present in the front legs. We also show that flower mites use electrostatic attraction to bridge the gap to the beaks of hummingbirds, for a few milliseconds becoming one of the fastest terrestrial organisms. Our results confirm that hummingbird flower mites evolved an additional sensory modality - electroreception - to quickly detect hummingbirds and use electrostatics to facilitate transportation onto their hosts.

Modeling spatiotemporal dynamics of Amblyomma americanum questing activity in the central Great Plains.

Ticks represent important vectors of a number of bacterial and viral disease agents, owing to their hematophagous nature and their questing behavior (the process in which they seek new hosts). Questing activity is notably seasonal with spatiotemporal dynamics that needs to be understood in detail as part of mediating and mitigating tick-borne disease risk. Models of the geography of tick questing activity developed to date, however, have ignored the temporal dimensions of that behavior; more fundamentally, they have often not considered the sampling underlying available occurrence data. Here, we have addressed these shortfalls for Amblyomma americanum, the most commonly encountered tick in the central Great Plains, via (1) detailed, longitudinal sampling to characterize the spatiotemporal dimensions of tick questing activity; (2) randomization tests to establish in which environmental dimensions a species is manifesting selective use; and (3) modeling methods that include both presence data and absence data, taking fullest advantage of the information available in the data resource. The outcome was a detailed picture of geographic and temporal variation in suitability for the species through the two-year course of this study. Such models that take full advantage of available information will be crucial in understanding the risk of tick-borne disease into the future.

Metabolic rate does not scale with body size or activity in some tick species.

Respiration in ticks is highly efficient and exceptionally low. Ticks can survive years between bloodmeals by having low activity and respiration to conserve energetic resources. Our objective was to compare metabolic (VCO2) and activity rates across 6 tick species. We predicted that VCO2 would be different among species and scale linearly with activity and body mass. Activity and CO2 production were measured for 32h in 6 tick species: Dermacentor andersoni, D. variabilis, Haemaphysalis longicornis, Rhipicephalus appendiculatus, R. microplus, and R. sanguineus. Individual ticks were measured for 30min three times to ensure breathing occurred. Absolute and mass-specific VCO2, total activity, body mass, and ventilation patterns were compared among species. As expected, ticks did not always breathe during the 30-minute measurements, especially R. sanguineus. Ventilation patterns differed among species with R. microplus having primarily cyclic patterns and R. appendiculatus having discontinuous gas exchange. VCO2 did not scale with body mass in most species. Haemaphysalis longicornis and R. sanguineus had the lowest VCO2; however, H. longicornis was the second most active species. Life history, including questing behavior and range expansion, could be contributing to differences between species. For instance, H. longicornis had exceptionally low metabolic rates despite above average activity levels, suggesting an energetic advantage which may underlie recently documented range expansions in North America. Our results demonstrate how ticks utilize energetic resources to maximize longevity. Future research describing questing behavior and distribution modeling may help explain differences in metabolic rates and activity and impacts on life history traits.

Climatic stress decreases tick survival but increases rate of host‐seeking behavior

AbstractTicks are vectors of many diseases and are expanding in geographic distribution. However, how ticks will fare in their new environments, where they may experience stressful climatic conditions at the expansion front, remains unclear. Since there is a trade‐off in ticks between behaviors that promote longevity and behaviors that promote reproduction, we hypothesized that extreme climatic stress reduces the survivorship of ticks but increases the frequency of tick host‐seeking behavior, or questing. Here, we used a novel method to simulate climatic stress on individual ticks of three species—Amblyomma americanum, Dermacentor variabilis, and Ixodes scapularis—to evaluate their survival, physiology, and questing behavior. The first experiment involved placing 144 adult ticks of each species in two temperature ranges (15–25°C and 25–35°C) and three relative humidity (RH) treatments (32%, 58%, and 84% RH). We assessed the ticks daily for survivorship and questing, and we measured water loss by comparing the mass of each tick when it died to when it was fully hydrated. In this first experiment, ticks in warmer and less humid conditions generally died faster than those in cooler and more humid conditions. Ticks of all three species were more likely to quest shortly before their death and consistently died after losing approximately 50%–56% of their total body water content, but Ixodes reached that threshold much faster than the other two species. The second experiment involved placing 18 ticks of each species at 35°C and 32% RH. We assessed the ticks every 3 h for survivorship, questing, and water loss. Ticks again were more likely to quest shortly before their death. With frequent checks, we were able to measure the dehydration tolerance more accurately and the rate of water loss. Ticks of all three species consistently died after losing approximately 51% of their total body water content. However, Ixodes lost water approximately 5 times faster than Amblyomma and 11 times faster than Dermacentor. These results demonstrate that severe climatic stress tilts the trade‐off toward higher questing rates but not higher overall questing time because of reduced survival rates.

Spatial repellents transfluthrin and metofluthrin affect the behavior of Dermacentor variabilis, Amblyomma americanum, and Ixodes scapularis in an in vitro vertical climb assay.

Repellents serve an important role in bite protection. Tick repellents largely rely on biomechanisms that induce responses with direct contact, but synthetic pyrethroids used as spatial repellents against insects have received recent attention for potential use in tick protection systems. An in vitro vertical climb assay was designed to assess spatial repellency against Dermacentor variabilis, Amblyomma americanum, and Ixodes scapularis adult, female ticks. Climbing behavior was assessed with and without the presence of two spatial repellents, transfluthrin and metofluthrin. Repellency parameters were defined to simulate the natural questing behavior of ambushing ticks, including measures of detachment, pseudo-questing duration, climbing deterrence, and activity. Significant effects were observed within each parameter. D. variabilis showed the greatest general susceptibility to each repellent, followed by A. americanum, and I. scapularis. The most important and integrative measure of repellency was climbing deterrence-a measure of the spatial repellent's ability to disrupt a tick's natural propensity to climb. Transfluthrin deterred 75% of D. variabilis, 67% of A. americanum, and 50% of I. scapularis. Metofluthrin was slightly more effective, deterring 81% of D. variabilis, 73% of A. americanum, and 72% of I. scapularis. The present study poses a novel paradigm for repellency and reports a preliminary assessment of spatial repellent effect on tick behavior. Further research will assess spatial repellency in a more natural setting, scale exposure conditions, and incorporate host cues.

A novel laboratory method to simulate climatic stress with successful application to experiments with medically relevant ticks.

Ticks are the most important vectors of zoonotic disease-causing pathogens in North America and Europe. Many tick species are expanding their geographic range. Although correlational evidence suggests that climate change is driving the range expansion of ticks, experimental evidence is necessary to develop a mechanistic understanding of ticks’ response to a range of climatic conditions. Previous experiments used simulated microclimates, but these protocols require hazardous salts or expensive laboratory equipment to manipulate humidity. We developed a novel, safe, stable, convenient, and economical method to isolate individual ticks and manipulate their microclimates. The protocol involves placing individual ticks in plastic tubes, and placing six tubes along with a commercial two-way humidity control pack in an airtight container. We successfully used this method to investigate how humidity affects survival and host-seeking (questing) behavior of three tick species: the lone star tick (Amblyomma americanum), American dog tick (Dermacentor variabilis), and black-legged tick (Ixodes scapularis). We placed 72 adult females of each species individually into plastic tubes and separated them into three experimental relative humidity (RH) treatments representing distinct climates: 32% RH, 58% RH, and 84% RH. We assessed the survival and questing behavior of each tick for 30 days. In all three species, survivorship significantly declined in drier conditions. Questing height was negatively associated with RH in Amblyomma, positively associated with RH in Dermacentor, and not associated with RH in Ixodes. The frequency of questing behavior increased significantly with drier conditions for Dermacentor but not for Amblyomma or Ixodes. This report demonstrates an effective method for assessing the viability and host-seeking behavior of tick vectors of zoonotic diseases under different climatic conditions.

Seasonal changes in questing efficiency of wild Amblyomma americanum (Acari: Ixodidae) nymphs

Understanding the factors which influence host-seeking behavior of ticks is essential to determine the risk they pose as a vector of pathogens. While many studies have evaluated the impact of environmental variables on tick behavior, few have examined how seasonal changes in physiological status may further modify patterns of activity. In this study, we measured differences in questing behavior of mid spring- and early summer-caught Amblyomma americanum nymphs held under standardized laboratory conditions. As both groups represent the same cohort of overwintered nymphs, we hypothesized that age-related changes in the older summer ticks may influence questing behavior. In each season, we collected nymphs from field and forest habitats in northeast Missouri, after which we placed each nymph individually in a desiccating vertical questing apparatus with a hydrating microenvironment at the base. On the day following collection, we recorded the height of each nymph in the apparatus bi-hourly from 04:00 to 22:00 and calculated the vertical displacement between consecutive observations. Despite displaying no differences in mean questing height throughout the experiment, active ticks collected in the summer group (n = 89) travelled greater cumulative distances between desiccating and hydrating microenvironments than those collected in the spring (n = 119). This suggests that questing efficiency decreases in summer nymphs to accommodate increased time allocation towards rehydration. While we observed no direct association between body size and distance travelled, body size of the nymphal population also decreased significantly from spring to summer. Overall, our results demonstrate that there are seasonal changes in how A. americanum responds to environmental conditions. To more accurately predict host-seeking behavior of ticks across seasons, models should incorporate physiological parameters of the active ticks in a given population.

Questing behavior of adult Amblyomma americanum (L.) in a laboratory setting</p >

Tick-borne diseases are on the rise globally; however, information is lacking about tick questing behavior. In this laboratory study, we explored tick preferences for stem type (plastic grass, wooden, and metal), questing height, and head orientation. Using 60 Amblyomma americanum adults over three 72-hour replicates, we determined that 21.7% of ticks quested at any given time and that ticks exhibited a strong preference to quest with their heads oriented downwards, irrespective of stem type. Individual ticks tended to quest on only one stem in this study, and a maximum of three. Nonetheless, ticks appeared to prefer questing on wooden and plastic grass stems over metal stems. We did not find an effect of time of day on tick questing rates. Increased understanding of tick questing behavior can improve vector control efforts.

Molecular identification of Haemaphysalis sulcata (Acari: Ixodidae) larval stages collected using the Berlese funnel in Northern Iran

Molecular detection techniques based on conventional PCR and sequencing of the partial genome of living organisms are considered an important tool for identification. BLASTn and comparison of evolutionary relationships between taxa are the next analysis steps on sequences. The construction of a phylogenetic tree is done using genetic distance difference; nucleotide substitution models and comparison with the outgroup. In the present study, two samples of tick larvae were collected in Mirafzal forest area (36°07'39.0N 53°35'45.0E) located in Mazandaran province, northern Iran using the Berlese funnel. Tick samples were initially identified using a generic identification key (Bregetova et al. 1955) and in order to determine specific identification of species, PCR of partial internal transcribed spacer 2 (ITS2) gene and Sanger sequencing were performed. Morphological results showed that tick samples belonged to Haemaphysalis genus because they had a relatively short capitulum, no eyes with festoons. DNA extraction, PCR amplification and sequencing of a partial fragment (ca. 1400-bp) ITS2 gene of tick sample were done, successfully. The results of BLASTn showed that the larvae belonged to the species Haemaphysalis sulcata. A nucleotide DNA sequence of ITS2 was submitted to GenBank under the accession number MW929218. The molecular technique of this study is recommended to identify immature and dead tick samples that cannot be reared until the adult stage. The present study was the first report on the identification of H. sulcata larvae isolated from the Berlese funnel. The result of this study will help to better understand the biology of this tick and the presence of immature larval stages with questing behaviour in the soil environment.

Cold hardening improves larval tick questing under low temperatures at the expense of longevity

Questing in ticks is essential for locating a host, and this behavioral response can occur at regionally specific low temperatures for most tick species. Little is known about the dynamics between tick questing behavior and temperature in ticks, specifically how this may impact other aspects of tick biology. Here, we examine whether cold hardening increases questing in three larval tick species (Ixodes uriae, Dermacentor variabilis, and Amblyomma americanum) at low temperatures and whether cold hardening impacts longevity. Rapid cold hardening and prolonged cold acclimation benefitted ticks by decreasing the temperature of chill coma onset, and increased survival, activity, and questing in ticks at low temperatures. Oxygen consumption increased at low temperatures following acclimation in larvae, suggesting this process has a distinct metabolic expense. This increased metabolism associated with hardening led to a substantial reduction in larval longevity as nutrient reserves are limited and cannot be replenished until a host is located. These studies suggest that tick larvae, and likely other developmental stages, require a delicate balance between the need for questing at low temperatures and survival until the first blood meal.

Impact of abiotic factors, habitat type and urban wildlife on the ecology of hard ticks (Acari: Ixodidae) in urban and peri-urban habitats

BackgroundTicks are increasingly acknowledged as significant vectors for a wide array of pathogens in urban environments with reports of abundant tick populations in recreational areas. The study aims to contribute to a better knowledge of the abiotic and biotic factors which impact the ecology of hard ticks in urban and peri-urban habitats in Romania.MethodsQuesting ticks were collected by flagging in seven recreational locations, from four types of habitats in Cluj-Napoca, Romania: parks; gardens; a cemetery; and peri-urban forests. Hedgehogs, birds and micromammals were also sampled and searched for ticks, using standard methods (i.e. torch-based searches, ornithological mist nets, snap-traps, etc.), while vegetation was evaluated on surveyed areas. Data on questing ticks were converted to abundance indices. Moodʼs median tests were used to assess the relationship between the abiotic and biotic factors and the abundance of questing ticks.ResultsTwo species of questing ticks were found: Ixodes ricinus (96.8%) and Haemaphysalis punctata (3.2%). Ixodes ricinus was also the predominant engorged tick collected from urban wildlife. For I. ricinus the highest mean total abundance index/location (total no. of ticks/100 m2) was recorded in the urban gardens (3.79, 95% CI: ± 1.59) and parks (2.68, 95% CI: ± 0.75), whereas the lowest mean total abundance index was noted in the peri-urban forests (0.06, 95% CI: ± 0.03) and the urban cemetery (0.04, 95% CI: ± 0.02). The adults and nymphs of I. ricinus displayed a bimodal activity pattern, while the larvae showed a unimodal questing behaviour with an autumnal peak. Positive correlations were found between the mean total abundance index and the rise in the daily mean temperature and relative humidity, and between the global abundance of questing ticks and the presence of hedgehogs in the respective locations (P < 0.01).ConclusionsTicks were collected in all the recreational sites surveyed in Cluj-Napoca. Ixodes ricinus was the dominant questing and engorged tick species. Several abiotic and biotic factors shape the ecology of ticks in Cluj-Napoca city, with climate and the local presence of suitable hosts being the most important.

A comparative evaluation of northern and southern Ixodes scapularis questing height and hiding behaviour in the USA.

Ticks display a distinct type of host-seeking behaviour called questing. It has been proposed that the questing behaviour of Ixodes scapularis explains the geographic variation in Lyme disease (LD) risk in the eastern USA because the northern population has been shown to quest more often than the southern population. The height at which questing occurs is variable and this study aimed to characterize questing height for I. scapularis. Ticks were collected from a northern and southern state (i.e. Maryland and Texas) and bioassays were conducted. We report that nymphs from Texas quested at lower heights compared to nymphs from Maryland. In addition, only Texas nymphs exhibited a behaviour we call 'hiding behaviour'. These results may reflect the different composition of hosts between these two areas as the south has a higher abundance of lizards. In contrast, there was no significant difference in questing height between Maryland adults and Texas adults which was to be expected since adults are feeding on white-tailed deer in both locations. If all southern I. scapularis nymphs are questing at lower heights, this might make them less likely to come into contact with humans and this may be contributing to the geographical difference in LD prevalence.

Free-living ticks (Acari: Ixodidae) in the Iguaçu National Park, Brazil: Temporal dynamics and questing behavior on vegetation

Basic knowledge of species, ecological preferences and behavior of ticks in preserved rainforests are scanty. We herein describe species richness, seasonal activity, altitude influence and questing height of ticks along 3375 m of animal trails within the largest remain of inland Atlantic rainforest in Brazil, the Iguaçu National Park (INP). Altogether, 2954 ticks from nine tick species were collected; Amblyomma brasiliense Aragão (49.09% of the specimens), Amblyomma coelebs Neumann (29.08%), Amblyomma incisum Neumann (12.49%), Haemaphysalis juxtakochi Cooley (5.21%), Amblyomma ovale Koch (2.64%), Amblyomma longirostre Koch (0.20%), Ixodes fuscipes Fonseca (0.17%), Ixodes loricatus Neumann (0.07%) and Amblyomma dubitatum Neumann (0.03%) (Ixodida: Ixodidae). There was an evident seasonal influence on tick host-questing activity, with each tick stage prevailing in a season before the following one, and a general trend of one tick generation per year. Adult tick questing height mode on vegetation showed a correspondence between tick species and preferred host size and height. Adult ticks quested higher than immatures, but several larval clusters were found high enough to seek for large-sized hosts as well. Nymphal and adult tick questing height increased significantly at higher altitudes within the park. Amblyomma incisum and A. coelebs were more abundant at higher and lower altitudes, respectively. The knowledge on tick species richness, ecological preferences and behavior herein obtained provides baseline information to understand tick-borne disease epidemiology occurring at landscapes under a strong anthropogenic impact.

Environmental Drivers of Questing Activity of Juvenile Black-Legged Ticks (Acari: Ixodidae): Temperature, Desiccation Risk, and Diel Cycles.

Vector feeding behavior can have a profound influence on the transmission of vector-borne diseases. In the case of black-legged ticks, Ixodes scapularis Say, which vectors the agents of Lyme disease, babesiosis, and other pathogens, the timing and propensity of questing can determine which hosts are fed upon as well as the risk of contact with humans. Yet we know little about the controls and constraints on tick host-finding behavior under natural conditions. Ticks must balance the need to quest for blood meal hosts with the risk of desiccation, all on a fixed energy budget. Prior research, primarily in the laboratory, has shown that questing activity varies with conditions (e.g., temperature, relative humidity), light-dark cycles, and energy reserves, but the findings have been idiosyncratic and the dominant factor(s) in nature remains unknown. We measured questing activity of nymphs and larvae throughout the day and night and over several weeks in enclosures across a range of suitable tick habitats within a site in the Northeast. Activity of nymphs increased slightly during dawn and dusk, opposite of larvae, and declined slightly with air temperature and rain, but these patterns were weak and inconsistent among replicate sites. Rather it appears a fraction of ticks were questing most of the time, regardless of conditions. Our study suggests neither climatic conditions or light-dark cycles have appreciable influence on tick questing behavior.

Nymphal Ixodes scapularis questing behavior explains geographic variation in Lyme borreliosis risk in the eastern United States

Most people who contract Lyme borreliosis in the eastern United States (US) acquire infection from the bite of the nymphal life stage of the vector tick Ixodes scapularis, which is present in all eastern states. Yet <5% of Lyme borreliosis cases are reported from outside the north-central and northeastern US. Geographical differences in nymphal questing (i.e., host-seeking behavior) may be epidemiologically important in explaining this latitudinal gradient in Lyme borreliosis incidence. Using field enclosures and a ‘common garden’ experimental design at two field sites, we directly tested this hypothesis by observing above-litter questing of laboratory-raised nymphal I. scapularis whose parents were collected from 15 locations (= origins) across the species' range. Relative to southern nymphs from origins considered to be of low acarologic risk, northern nymphs from high-risk origins were eight times as likely to quest on or above the surface of the leaf litter. This regional variation in vector behavior (specifically, the propensity of southern nymphs to remain under leaf litter) was highly correlated with Lyme borreliosis incidence in nymphs’ counties of origin. We conclude that nymphal host-seeking behavior is a key factor contributing to the low incidence of Lyme borreliosis in southern states. Expansion of northern I. scapularis populations could lead to increased incidence in southern states of Lyme borreliosis and other diseases vectored by this tick, if the ‘northern’ host-seeking behavior of immigrant nymphs is retained. Systematic surveillance for I. scapularis nymphs questing above the leaf litter in southern states will help predict future geographic change in I. scapularis-borne disease risk.

Progressive behavioural, physiological and transcriptomic shifts over the course of prolonged starvation in ticks.

Ticks are obligatorily hematophagous but spend the majority of their lives off host in an unfed state where they must resist starvation between bouts of blood feeding. Survival during these extended off-host periods is critical to the success of these arthropods as vectors of disease; however, little is known about the underlying physiological and molecular mechanisms of starvation tolerance in ticks. We examined the bioenergetic, transcriptomic and behavioural changes of female American dog ticks, Dermacentor variabilis, throughout starvation (up to nine months post-bloodmeal). As starvation progressed, ticks utilized glycogen and lipid, and later protein as energy reserves with proteolysis and autophagy facilitating the mobilization of endogenous nutrients. The metabolic rate of the ticks was expectedly low, but showed a slight increase as starvation progressed possibly reflecting the upregulation of several energetically costly processes such as transcription/translation and/or increases in host-seeking behaviours. Starved ticks had higher activity levels, increased questing behaviour and augmented expression of genes related to chemosensing, immunity and salivary gland proteins. The shifts in gene expression and associated behavioural and physiological processes are critical to allowing these parasites to exploit their ecological niche as extreme sit-and-wait parasites. The overall responses of ticks to starvation were similar to other blood-feeding arthropods, but we identified unique responses that could have epidemiological and ecological significance for ticks as ectoparasites that must be tolerant of sporadic feeding.

Uranotaenia unguiculata Edwards, 1913 are attracted to sound, feed on amphibians, and are infected with multiple viruses

BackgroundUranotaenia unguiculata Edwards, 1913 is a species of mosquito (Diptera: Culicidae) native to central Europe. Recently a novel lineage of the West Nile virus (WNV-lineage 4c) was identified in pools of adult female Ur. unguiculata. To increase the body of knowledge about this species, various trapping methods were evaluated to determine the most efficient method for capturing adult female Ur. unguiculata.ResultsSound traps collected equivalent numbers of female Ur. unguiculata as low-hanging light-baited downdraft traps. Hosts were identified as Pelophylax lessonae and P. ridibunda (Anura: Ranidae) species group frogs from the blood found in engorged females. In addition to confirming infection by WNV-lin. 4c, a potentially integrated flavivirus sequence was detected in male mosquitoes. A novel Alphamesonivirus 1 (Nidovirales: Mesoniviridae) was found to be widespread in the Ur. unguiculata population and is herein described.ConclusionsEfficient collection methods for Ur. unguiculata for arbovirus surveillance reflect mosquito questing behavior. Uranotaenia unguiculata targets frog species which call from the water, and it is likely that the novel WNV-lin. 4c is maintained in a frog-mosquito transmission cycle. The improved trapping methods listed here will assist future studies of the vector status of Ur. unguiculata for WNV and other arboviruses.

Behavioral asymmetries in ticks – Lateralized questing of Ixodes ricinus to a mechatronic apparatus delivering host-borne cues

Ticks are considered among the most dangerous arthropod vectors of disease agents to both humans and animals worldwide. Lateralization contributes to biological fitness in many animals, conferring important functional advantages, therefore studying its role in tick perception would critically improve our knowledge about their host-seeking behavior. In this research, we evaluated if Ixodes ricinus (L.) (Ixodiidae) ticks have a preference in using the right or the left foreleg to climb on a host. We developed a mechatronic device moving a tuft of fox skin with fur as host-mimicking combination of cues. This engineered approach allows to display a realistic combination of both visual and olfactory host-borne stimuli, which is prolonged over the time and standardized for each replicate. In the first experiment, the mechatronic apparatus delivered host-borne cues frontally, to evaluate the leg preference during questing as response to a symmetrical stimulus. In the second experiment, host-borne cues were provided laterally, in an equal proportion to the left and to the right of the tick, to investigate if the host direction affected the questing behavior. In both experiments, the large majority of the tested ticks showed individual-level left-biased questing acts, if compared to the ticks showing right-biased ones. Furthermore, population-level left-biased questing responses were observed post-exposure to host-mimicking cues provided frontally or laterally to the tick. Overall, this is the first report on behavioral asymmetries in ticks of medical and veterinary importance. Moreover, the mechatronic apparatus developed in this research can be exploited to evaluate the impact of repellents on tick questing in highly reproducible standardized conditions.

Ambush behavior of the tick Amblyomma sculptum (Amblyomma cajennense complex) (Acari: Ixodidae) in the Brazilian Pantanal

We herein describe the ambush behavior of Amblyomma sculptum (Berlese 1888), a widespread and epidemiologically important tick in Brazil. Along two years of sampling by visual search in the Brazilian Pantanal, A. sculptum ticks were observed on the vegetation and in the leaf litter. Most of the ticks were observed between 10 and 50cm above ground level and less than five percent of the total were positioned below 10cm, indicating that they are seeking for middle or large-sized hosts. In both seasons, vapor saturation deficit was low during the morning. No significant relationship was found between questing ticks and daytime interval of observation or saturation deficit. However, questing tick numbers seem be higher in the end of the morning, when saturation deficit reaches its peak. Behavioral patterns of A. sculptum ticks observed in Pantanal underscore the occurrence of this tick and human contact at green anthropogenic sites. Considering A. sculptum questing behavior, inferences on human behavioral patterns that enhance or avoid contact with ticks are discussed.

Larval behaviour of the winter tick, Dermacentor albipictus (Acari:Ixodidae): evaluation of CO2 (dry ice), and short- and long-range attractants by bioassay

ABSTRACTWe report that larvae of Dermacentor albipictus can detect CO2 from a dry ice source at distances up to 3.0 m and initiate increased activation, movement and questing behaviour. The maximum distance that larvae spread was ~0.5 m after 24 h, compared to <0.1 m without CO2 as an excitant. In Petri dish bioassays, 2,6-dichlorophenol, methyl salicylate and o-nitrophenol acted as attractants, albeit with mixed results at 1.0 m, 2.0 m and 3.0 m, respectively. The 2,6-dichlorophenol target was the most effective, although few larvae crawled >1.0 m and CO2 was required as a carrier. In Petri dish bioassays, larvae arrested into dense aggregations on eggshells collected after hatching. This arrestment was associated with the eggshell lipids cholesteryl oleate and cholesteryl palmitate, implying that cholesteryl esters act as assembly pheromones for larvae. Faster response was evident at 65% relative humidity (RH) versus 95% RH. Larvae were not arrested by tick excreta, guanine or related purines. Although larvae can be activated from long distances, in the field they remain fairly localized around the egg mass (hatching location), have limited dispersal and are more responsive in drier conditions. Given their arrestant properties, cholesteryl oleate and cholesteryl palmitate may prove useful in pheromone-assisted control methods because D. albipictus larvae are the single stage of infestation to the host.