Different Populations of Blacklegged Tick Nymphs Exhibit Differences in Questing Behavior That Have Implications for Human Lyme Disease Risk

Animal behavior can have profound effects on pathogen transmission and disease incidence. We studied the questing (= host-seeking) behavior of blacklegged tick (Ixodes scapularis) nymphs, which are the primary vectors of Lyme disease in the eastern United States. Lyme disease is common in northern but not in southern regions, and prior ecological studies have found that standard methods used to collect host-seeking nymphs in northern regions are unsuccessful in the south. This led us to hypothesize that there are behavior differences between northern and southern nymphs that alter how readily they are collected, and how likely they are to transmit the etiological agent of Lyme disease to humans. To examine this question, we compared the questing behavior of I. scapularis nymphs originating from one northern (Lyme disease endemic) and two southern (non-endemic) US regions at field sites in Wisconsin, Rhode Island, Tennessee, and Florida. Laboratory-raised uninfected nymphs were monitored in circular 0.2 m² arenas containing wooden dowels (mimicking stems of understory vegetation) for 10 (2011) and 19 (2012) weeks. The probability of observing nymphs questing on these stems (2011), and on stems, on top of leaf litter, and on arena walls (2012) was much greater for northern than for southern origin ticks in both years and at all field sites (19.5 times greater in 2011; 3.6–11.6 times greater in 2012). Our findings suggest that southern origin I. scapularis nymphs rarely emerge from the leaf litter, and consequently are unlikely to contact passing humans. We propose that this difference in questing behavior accounts for observed geographic differences in the efficacy of the standard sampling techniques used to collect questing nymphs. These findings also support our hypothesis that very low Lyme disease incidence in southern states is, in part, a consequence of the type of host-seeking behavior exhibited by southern populations of the key Lyme disease vector.     

A test of the relationship between cuticular melanism and immune function in wild‐caught Mormon crickets

Cuticular melanism and innate immune parameters can share common physiological pathways in insects, and this functional connection may contribute to the maintenance of insect colour polymorphisms. However, evidence linking colouration and immune function has been equivocal, particularly when tested in wild populations. The present study investigates phenotypic links between colouration and immune function in migratory Mormon crickets (Anabrus simplex, Haldeman), in which juveniles occur in conspicuous colour variants but mature to become uniformly melanic adults. Wild‐caught insects are used to evaluate the relationship between juvenile colouration and three immune parameters: encapsulation ability, lysozyme‐like activity and phenoloxidase activity. As nymphs, brown crickets are better able to encapsulate an inert implant introduced into the haemocoel than green crickets, although the difference is slight and ceases after they all become darkly‐coloured adults. By contrast, adults that develop from brown nymphs have a higher basal phenoloxidase activity than those that develop from green nymphs, regardless of the fact that all adults are brown. Intrinsic factors other than colouration exert larger effects on immunity: males show stronger encapsulation responses but lower phenoloxidase activity than females, suggesting a sex‐specific trade‐off between these two immune parameters, and adults exhibit higher immune function than nymphs. In summary, modest support is found for a correlation between cuticular melanism and increased immune function in wild Mormon crickets. Additional intrinsic factors such as developmental stage and sex appear to interact with colouration and have a more substantial connection to immune function in the wild.     

Responses of Second-Instar Male Nymphs of Four Mealybug Species (Hemiptera: Pseudococcidae) to Conspecific and Heterospecific Female Sex Pheromones

The response of the late second-instar male nymphs of the mealybug species (Hemiptera, Pseudococcidae), Planococcus citri (Risso), Planococcus ficus (Signoret), Pseudococcus cryptus Hempel Nipaecoccus viridis (Newstead), to their conspecific and heterospecific female pheromone was studied. Males that exhibited the typical appearance of late-second-instar nymphs were tested. The male behavior was monitored soon after their exposure to the tested female sex pheromone in glass Petri dish arenas. Male nymph behavior toward the pheromone source was characterized based on their aggregation on the disks in the arena. Males of all four tested mealybug species were attracted to their conspecific female pheromone. By contrast, almost no interceptions of male nymphs with disks impregnated with a heterospecific female pheromone were observed. The mode of attraction of each of male nymphs of P. ficus, among most of the tested individuals (>80%), to the conspecific female sex pheromone, (S)-lavandulyl senecioate and or (S)-lavandulyl isovalerate, was the same as the mode of attraction latter on as adult. We suggest that by being attracted to the conspecific pheromone these males may direct themselves to a suitable pupation site near conspecific non-sibling mature females, thus preventing inbreeding. The repellency of heterospecific sex pheromone to males that are looking for a pupation site suggests that the latter try to avoid close contact with heterospecific females.     

Sex differentiation studies relating to releaser aggregation pheromones of the desert locust,Schistocerca gregaria

Behavioural responses of nymphs and adults in the gregarious phase of the desert locust,Schistocerca gregaria (Forskal) (Orthoptera: Acrididae) were investigated in a single-chamber bioassay system to a choice of two columns of air, one permeated with airborne volatiles emanating from either sex of nymphs or adults and the other untreated. There was no sexual differentiation in the production of or response to nymphal volatiles. Young adults of either sex did not produce a stimulus with significant activity. Of the older adults, only the males produced the aggregation stimulus to which both sexes were equally responsive. Charcoal-trapped volatiles from the two sexes of nymphs and adults evoked similar aggregation responses. Antennae of the older adults showed significantly higher EAG responses than those of fifth instar nymphs to all four volatile collections, of which volatiles from older adult males were the most stimulatory and evoked the highest EAG amplitudes.     

Mapping of spatiotemporal distribution of Tibraca limbativentris Stal (Hem.: Pentatomidae) in flooded rice crop in Southern Brazil

The rice stem bug, Tibraca limbativentris Stal (Hem.: Pentatomidae), is one of the main insect pests in Brazilian rice crops. Knowledge of its spatiotemporal distribution can support the development of an effective sampling system and improve IPM strategies. This study aimed to map the spatiotemporal distribution of rice stem bug in flooded rice crop in Southern Brazil. Flooded rice crop was scouted for two seasons to estimate insect densities. Four occurrence categories were observed: no insect, only adults, only nymphs, and both phases. The relationship between phenological stages and insect categories was established. Interpolation (mapping) of occurrence categories of T. limbativentris was performed by multiquadric equations. In two seasons during the rice cycle, the results indicate that rice stem bug “adults” were the most abundant category until the mid-vegetative phase of the rice; “nymphs” were the most abundant from the end of vegetative phase; “adults + nymphs” occurred from the beginning of reproductive phase; there were no rice stem bugs in more than 66% of the area, as they were most concentrated near the edge of the crop. The information presented here provides further knowledge about T. limbativentris spatiotemporal dynamics that can be applied to improve IPM strategies, such as developing sampling plans and localized control measures at the edge of rice fields.     

The role of direct chilling injury and inoculative freezing in cold tolerance of Amblyomma americanum, Dermacentor variabilis and Ixodes scapularis

Abstract. Supercooling points and chill tolerance were compared among nymphs and adults of the ixodid ticks Dermacentor variabilis, Amblyomma americanum and Ixodes scapularis (Acari: Ixodidae).Supercooling points in the range of <-22 to -18°C were observed for nymphs, and -22 to -8°C for adults.The lower lethal temperatures observed under dry conditions, -14 to -10°C, were warmer than the supercooling points, but still much colder than -4.8°C, the lowest temperature recorded from a likely tick habitat in southwestern Ohio.Based on our experiments, spontaneous freezing and direct chilling injury are not significant mortality factors in these species in the field.Mortality was observed between -5 and -3°C for A.americanum and D.variabilis nymphs chilled for 2 h while in direct contact with ice.This mortality is probably due to inoculative freezing.Given the requirement for a rather humid microhabitat for off-host survival, these findings suggest that inoculative freezing is an important cause of overwintering mortality in these medically important species.     

Establishment potential of the predatory mirid <Emphasis Type="Italic">Dicyphus hesperus</Emphasis> in northern Europe

The predatory mirid Dicyphus hesperus Knight (Hemiptera: Miridae) is native to North America. The species has been used for the control of glasshouse whitefly on aubergine in the Netherlands, and is currently being evaluated for continued and wider release in Europe. Field and laboratory studies were conducted on a population collected from southern California, USA, to assess the cold tolerance and potential for outdoor establishment under prevailing northern European climates. The supercooling points (whole animal freezing temperatures) of nymphal and adult insects were around −20°C. The lethal temperatures (LTemp₅₀) of non-diapausing nymphs and adults and diapausing adults were close to their respective freezing temperatures at −17.6, −17.6 and −19.2°C. At 5°C, the LTime₅₀ was 54, 101.7 and 117.5 days for fed nymphs, non-diapausing and diapausing adults respectively. When first instar nymphs were placed in the field in winter, starved samples died out after 70 days, but 5% of the fed nymphs survived until the end of winter (140 days) and developed to adult on return to the laboratory. After a similar 5-month field exposure, 50% of fed diapausing adults and 15% of fed non-diapausing adults were still alive at the end of winter, whereas starved diapausing adults died after 140 days. On return to the laboratory after 5 months in the field, both diapausing and non-diapausing adults mated and laid eggs, forming viable populations. Overall, the field and laboratory experiments indicate that this population of D. hesperus is able to enter diapause and that winter temperatures are not a barrier to establishment in northern Europe.     

Territorial behavior of the Korean wood-feeding cockroach, <Emphasis Type="Italic">Cryptocercus kyebangensis</Emphasis>

We investigated the territorial behavior of Cryptocercus kyebangensis from the viewpoint of offspring protection. Aggressive behavior against adult intruders occurred more frequently in resident adults with their young than those without their young. Resident adults without offspring more frequently attacked intruders of the same sex than those of the opposite sex, whereas those with offspring attacked regardless of the sex of adult intruders. In paired residents, cooperative territorial behavior occurred in a higher proportion in pairs with offspring than those without offspring. Responses of residents to nymphal intruders depended on the head capsule size of the intruders. Resident adults rarely attacked intruders smaller than their own offspring but frequently attacked larger intruders. Resident nymphs frequently attacked intruders smaller than themselves, whereas they escaped or showed jerking behavior when they encountered larger intruders. Electronic Publication     

Acclimatisation in the green spruce aphid, Elatobium abietinum

Thin layer chromatography separation of 80% ethanol extracts of adult Elatobium abietinum revealed the presence of the polyhydric alcohol mannitol in aphids overwintering outdoors but not in aphids kept permanently indoors at 15°C. After 3 days at 15°C no traces of mannitol were left in overwintering aphids. Mean freezing temperatures of outdoor, unfed instar I nymphs were about 4°C lower than those of unfed instar I nymphs produced at 15°C. Mean freezing temperatures of overwintering adults were considerably higher than those of unfed instar I nymphs and showed no changes associated with time at 15°C following transference indoors. Similarly, mean freezing temperatures of Sitka spruce needles transferred to 15°C did not change. It was concluded that, although freezing was mainly avoided by supercooling, the presence of mannitol lowered the true freezing temperature of aphid haemolymph and, consequently, the actual freezing temperatures of nymphs produced under cold conditions. However, the considerable increase in freezing point temperatures caused by imbibition of plant sap masked these acclimatisation changes in feeding aphids.     

Alarm pheromone eliciting attack and escape responses in the sugar cane woolly aphid,Ceratovacuna lanigera (Homoptera,Pemphigidae)

All nymphal stages and apterous adults ofCeratovacuna lanigera secreted droplets containing an alarm pheromone from their abdominal cornicles when stimulated with a pin. When fresh droplets on piece of filter paper were put near the aphids, 1st-instar nymphs originating from apterous adults attacked it, but the advanced instar nymphs and adults escaped from the spot. Nymphs and adults originating from alate adults showed escaping behavior only. These different responses strongly correlated with morphological differences, such as relative lengths of the frontal horns and legs. When a syrphid larva,Allograpta javana, was placed on a colony of aphids, many 1st-instar nymphs attacked it using their frontal horns. During the attack, these nymphs spontaneously secreted droplets and adhered them to the syrphid body. When these droplets were placed on the body, the syrphid larva was agressively attacked by additional 1st-instar nymphs.     

Morphology of Hydrozetes confervae () and H. parisiensis (Acari: Oribatida: Hydrozetidae), and keys to European species of Hydrozetes Berlese, 1902

Hydrozetes confervae [Schrank, F.P., 1781. Enumeratio insectorum Austriae indigenorum. Augustae Vindeliciorum, 1-552] and Hydrozetes parisiensis [Grandjean, F., 1948. Sur les Hydrozetes (Acariens) de l’Europe occidentale. Bull. Mus. Nat. Hist. nat., Paris, ser. 2(20), 328-335] are common European species of the aquatic oribatid mite family Hydrozetidae, but their external morphology is incompletely known. We describe and illustrate their larvae for the first time, and the nymphs of both species are redescribed and illustrated. Juveniles of the two species differ mainly by the position of gastronotal seta lm in relation to the opisthonotal gland opening (gla) and by the number of extremely long posterior hysterosomal setae in nymphs. In H. confervae seta lm inserts anteromedial to gla, but posterior to gla in H. parisiensis; nymphs of H. confervae have three pairs of long hysterosomal setae, while H. parisiensis has a larger, but variable number. Separate identification keys for the larvae, nymphs and adults of all known European species of Hydrozetes Berlese, 1902, are presented.     

A miniaturized assay to quantify effects of chemicals or physical stimuli upon locust activity

Solitary and gregarious locusts differ in many traits, such as body color, morphometrics and behavior. With respect to behavior, solitary animals shun each other, while gregarious animals seek each other＇s company, hence their crowding behavior. General activity, depending on the temperature, occurs throughout the day but is much lower in solitary locusts. Solitary locusts occasionally fly by night while gregarious locusts fly regularly during daytime （swarming）. In search of new assays to identify substances that control or modify aspects of （phase） behavior, we designed a simple activity assay, meant to complement existing behavioral measurement tools. The general activity is reflected in the number of wall hits, that is, the number of contacts between the locust and the vertical walls of a small arena. Using this single parameter we were able to quantify differences in total activity of both nymphs and adults of isolation-reared （solitary）, regrouped- and crowdreared （gregarious） locusts under different conditions. Furthermore, we demonstrate that there are inter- and intra-phase dependent differences in activities of 5th instar nymphs afar injections of the three different adipokinetic hormones.     

Orientation Behavior with and Without Visual Cues in Newly Hatched and Adult Praying Mantis

In the praying mantis, vision plays a major role even in newly hatched nymphs, which have compound eyes that are not yet fully developed. This study examines how this factor affects the visual orientation behavior of freely mobile Mantis religiosa. Mantises from three age groups (nymphs newly hatched to 2 h old, three-day-old nymphs, and three- to four-month-old female and male adults) were placed in a completely unprotected open area, either with or without visual cues in the surroundings. As a visual cue, five vertical rods with high-, medium- or low-luminance contrast, having a vertical extension of 45° and overall horizontal extension of 40°, were presented at a distance of 300 mm, simulating a group of plant stems with differing contrasts. The mantis search behavior, probability of reaction to the visual cues, distance at which the first target-related reaction occurred, and target approach behavior were investigated. It was found that the search behavior differed in the different age groups. With high-contrast visual cues, newly hatched nymphs performed similarly to adults, but this was not the case for medium- and low-contrast visual cues. Visual performance was greatly improved 3 days after hatching, presumably due to the complete hardening of the cuticle. Nevertheless, despite differences in visual acuity, it was found that even newly hatched nymphs used visual orientation mechanisms similar to those of adult mantises, including fixation, scanning and peering.     

Factors affecting the cold hardiness of the peach-potato aphid Myzus persicae

Supercooling point studies were used to investigate the factors influencing the cold hardiness of the peach-potato aphid Myzus persicae, a freezing-susceptible insect. Overwintering adults lost cold hardiness as winter progressed, with a variable proportion showing a marked reduction in supercooling ability. Cold hardiness increased in spring so that all individuals demonstrated extensive supercooling ability typical of aphids reared in the laboratory at 20°C with a long photoperiod; these levels of cold hardiness were maintained in the field during summer and early autumn. First instar nymphs demonstrated considerable cold hardiness all year. Surface moisture caused inoculative freezing in some first instar nymphs and adults when supercooled, but the majority were unaffected. In the laboratory, adults starved for 7 days at 5°C showed distinct losses of supercooling potential equivalent to those observed in the field during mid to late winter. No loss of cold hardiness was found in first instar nymphs starved under the same conditions. The results demonstrate that the cold hardiness characteristics of M. persicae are atypical of those observed in other freezing-susceptible insects and it is suggested that continued feeding during mild winter conditions allows maintenance of cold hardiness particularly in adult aphids, and provides a possible explanation for the successful anholocyclic overwintering of M. persicae during such winters.     

Morphology and distribution of antennal sensilla of the tarnished plant bug,Lygus lineolaris (Palisot de beauvois) (Hemiptera: Miridae)

Sensilla on the antennae of adult and last-instar nymphs of the tarnished plant bug, Lygus lineolaris (Hemiptera: Miridae), were examined with light, scanning and transmission electron microscopy. Six different types were identified in adult females and males and 5 types in last-instar nymphs: types 1 and 4 are sensilla trichodea, 2 and 3 are sensilla chaetica, and 5 and 6 are sensilla basiconica. Type 1 are located at distal region of terminal segment and type 2 are located at distal regions of proximal 3 segments in both adults and nymphs. Type 3 is present on all segments, more numerous on scape and pedicel and less abundant on distal third and fourth segments in both adult and nymphal stages. Types 4 and 6 are absent on the scape and present on the distal 3 antennal segments in adults, but they are present only on the distal-most antennal segment in nymphs. Type 5 sensilla are present only on second antennal segments in adults and are absent in nymphs. Sexual dimorphism is observed in total numbers: there are significantly more type(s) 3, 4, 5 and 6 sensilla in adult males than adult females. Types 1, 4 and 5 are multiporous with thin cuticle, branched dendrites and pore tubules which suggests an olfactory function. These sensilla have 3, 3 and 2 neurons, respectively. The type 6 sensillum has an apical pore and pores in the cuticular wall, and is innervated by 5 nerve cells with unbranched dendrites. Sensillar types 2 and 3 have thick cuticle, a single apical pore and nerve cells with unbranched dendrites. Type 2 has 1 neuron and type 3 has 2 chambers and 2 nerve cells.     

Natural infestation of Hydrochoerus hydrochaeris by Amblyomma dubitatum ticks

Natural infestation of Amblyomma dubitatum in relation to individual specific attributes of Hydrochoerus hydrochaeris such as sex, body mass and body condition was analyzed. The anatomical distribution of A. dubitatum on H. hyrochaeris was also evaluated. Prevalence of adults and nymphs were significantly higher than prevalence of larvae. Non-significant differences in the infestation levels were found among host sex. Multiple regression analysis did not show any statistically significant association among the level of infestation with ticks and body mass and body condition of the host. All parasitic tick stages were collected in all five anatomical areas of the host, but they exhibited significant differences in feeding site preference. Factors associated to the host which determine the high levels of infestation with A. dubitatum could be assigned to a combination of population-level properties of the host as abundance, ubiquity and aggregation, rather than individual specific attributes related to body condition, body mass or sex.     

Phenology and demography of Homalodisca coagulata (Hemiptera: Cicadellidae) in southern California citrus and implications for management

Populations of Homalodisca coagulata (Say) were sampled from citrus orchards in southern California, USA to characterize and quantify seasonal occurrences of nymphs and adults with the goal of identifying management opportunities through well-timed treatments and/or natural enemy releases. Higher densities of H. coagulata in 2001 contributed to a complete seasonal profile that began in early spring with the emergence of first instar nymphs and their progression through five nymphal instars lasting until mid-August. Adult emergence began in mid-June with peak adult densities attained from mid to late August followed by a gradual decline through autumn. A persistent and significant male bias was observed in the adult sex ratio from the time of first emergence through mid-October in oranges; the same trend was present in lemons, but with more variability. Adult densities gradually declined through the winter months into the following spring before rapidly increasing again in June as the 2002 spring generation of nymphs began emerging as adults. The seasonal timing of nymphs and adults in 2002 was nearly identical to that observed the previous year. Phenology data from both years were incorporated into a stochastic, temperature-dependent model that predicts the occurrences of H. coagulata stages through time. Applications of imidacloprid early in the spring generation of nymphs proved very effective at reducing nymphs and sustaining lower densities of adults through summer.     

Prey Preference of Aphidoletes Aphidimyza on Acyrthosiphon Pisum: Effect of Prey Color and Size

Color polymorphism in insects as well as factors contributing to its occurrence and maintenance have been of a great interest. Pea aphid (A. pisum) exhibits a noteworthy color polymorphism which occurs as red and green. The preference of the predatory gall midge Aphidoletes aphidimyza (Rondani) for the two color morphs of pea aphid at two life stages (adult and second instar) was investigated. Red adults, red nymphs, green adults, and green nymphs were offered to the larvae of the predator in different sets. The larvae attacked red aphids more than green ones of a same size. But whenever adults were offered along with nymphs, the nymphs experienced significantly more attacks. Although visual cues could result in more predation on red aphids, our findings showed that the size of aphids was the more important factor. The results showed that previous findings, suggesting more predation on the red morph, are valid when the same size of aphids is used. The ovipositing females exhibited no differences in oviposition choice between stalks infested with either of two colors.     

Comparison of sampling methods for determining relative densities of Homalodisca vitripennis (Hemiptera: Cicadellidae) on citrus

Four sampling methods that included A-Vac, D-Vac, pole-bucket, and beat-net devices were evaluated for estimating relative densities of glassy-winged sharpshooter, Homalodisca vitri-pennis (Germar) (Hemiptera: Cicadellidae) nymphs and adults on citrus (Citrus spp.) trees. All four methods produced similar temporal and spatial distribution profiles, although significant differences in quantities of H. vitripennis adults and nymphs caught by each device were observed. The four sampling methods also showed a consistent male bias in adult populations across a range of densities, suggesting that previously reported male-biased sex ratios in H. vitripennis adult populations are real and not a product of sampling bias. A strong relationship (R2 = 0.95) between the monitoring methods we evaluated and yellow sticky trap catches of female H. vitripennis adults suggest that yellow sticky trap catches may provide a good relative index of infestation levels in citrus trees. Based on quantitative analyses examining precision and cost, the pole bucket was the most efficient method for sampling nymphs, and it was as efficient as the beat-net method for sampling adults and both stages combined. In addition to these analyses, consideration of other sampling characteristics such as added flexibility in sampling and higher sensitivity in detecting infestations within individual trees helped to fortify the conclusion that the pole bucket was the best overall sampling method of those tested.