Comparative Quantification of Trail-Following Behavior in Pest Ants.

A comparison of trail-following movement parameters of six major urban pest ants, Nylanderia fulva (Forel) (Hymenoptera: Formicidae), Pheidole megacephala, Linepithema humile (Mayr), Solenopsis invicta Buren, Paratrechina longicornis (Forel), and Technomyrmex albipes (Smith) demonstrated several differences in velocity of movement, amplitude of the deviations from a marked trail, percent fidelity to the trail, number of curves per unit of trail, and trail-following accuracy. Paratrechina longicornis and N. fulva had the largest deviations from the marked trails and moved three times faster (25-30 mm/s) along the trail than the slowest ant, S. invicta (< 10 mm/s), with other ants following between these extremes. Species differences in relation to going toward or returning from food were observed in a few cases, especially with Pa. longicornis for which velocity, amplitude, and trail fidelity differed between the foraging and return trails. Quantification of ant trail-following movement parameters can be useful in understanding the mechanics of ant movement and may be important in testing specific strategies and products that disrupt trail-following behavior.

Sticky Trap Design Considerations for Entrapping Bed Bugs.

Little evidence has been presented on the usefulness of sticky traps for monitoring bed bugs, Cimex lectularius. We examined how the surface roughness around the adhesive of a sticky trap affects both bed bug behavior and adhesive entrapment. In the first assay, bed bugs were placed onto acetate paper discs with different roughness averages (Ra). Each disc was surrounded by sticky trap adhesive and number of captured bed bugs were recorded. The second assay was set up similarly to the first assay except that the outer portion of the acetate disc had a different Ra than the center. In the third assay, bed bugs were placed into circular acetate arenas where they were surrounded by different Ra treatments. The number of times the bed bugs contacted the Ra treatment but did not cross onto the treatment was recorded. Results of these assays showed that as the acetate surfaces got smoother (lower Ra), bed bugs were more likely to get trapped in sticky trap adhesives but also less likely to travel across the smoother surfaces they encountered. A sticky trap design with a smooth plastic film around the adhesive was tested in the field to see if it could capture bed bugs in apartments with known bed bug activity. This trap was not only able to capture bed bugs but was also able to detect unknown German cockroach, Blattela germanica, infestations. Sticky trap designs with smooth surfaces around an adhesive could be used to monitor not only bed bugs but also German cockroaches.

Mosquitocidal Chips Containing the Insect Growth Regulator Pyriproxyfen for Control of Aedes aegypti (Diptera: Culicidae).

Aedes aegypti were exposed to water treated with mosquitocidal chips containing the insecticide pyriproxyfen in a polymer formulation. Chips were tested under different conditions; different water volumes, in containers made of different material, and in water with different levels of organic matter. Treated chips caused 100% mortality of Ae. aegypti during their pupal stage independent of size or type of container, and the mount of organic matter contained in the water to which the mosquito larvae were exposed. When mosquitocidal chips were used in >25% of the oviposition containers within their cages, there was a significant control of the mosquito populations. Mosquitocidal chips worked in different environments, caused significant mosquito population decreases, and were effective in controlling Ae. aegypti.

A Low-Cost, Passive Release Device for the Surveillance and Control of Mosquitoes.

Mosquitoes continue to be a major threat to global health, and the ability to reliably monitor, catch, and kill mosquitoes via passive traps is of great importance. Global, low-cost, and easy-to-use outdoor devices are needed to augment existing efforts in mosquito control that combat the spread of disease, such as Zika. Thus, we have developed a modular, portable, non-powered (passive), self-contained, and field-deployable device suitable for releasing volatiles with a wide range of applications such as attracting, repelling, and killing mosquitoes. This unique device relies on a novel nested wick and two-reservoir design that achieves a constant release of volatiles over several hundred hours. Devices loaded with one of either two compounds, geraniol or 1-methylpiperazine (MP), were tested in a controlled environment (32 °C and 70% relative humidity), and both compounds achieved a constant release from our devices at a rate of 2.4 mg/h and 47 mg/h, respectively. The liquid payload can be volatile attractants or repellants as well as mosquitocide-containing feeding solutions for capture and surveillance. This low-cost device can be utilized for both civilian and military mosquito control purposes, but it will be particularly important for protecting those in economically repressed environments, such as sub-Saharan Africa and Central and South America.

Laboratory Evaluation of a Novel Lethal Ovitrap for Control of Aedes aegypti.

Aedes aegypti (L.) (Diptera: Culicidae) are known to flourish in a variety of natural and residential habitats and are competent vectors of at least 22 different arboviruses, including dengue, chikungunya, and Zika. Their global distribution, anthropophilic nature, and vector competency make them species of interest for control. A novel durable dual-action lethal ovitrap (DDALO) with combined larviciding and adulticiding effects, as well as a slow-release polymer (isobutyl methacrylate), was designed to target Ae. aegypti. The use of the DDALO resulted in high adult mosquito mortality (~95-100%) in no-choice laboratory cage studies targeting gravid females and successfully prevented all deposited eggs from hatching. Aging of the traps caused some loss in activity over time, but they still caused adult mortality (~50%) and continued to prevent successful hatching of eggs for 6 mo. Oviposition preference studies resulted in ~4.5 times as many larvae developing in untreated DDALOs compared with the other containers combined. Small-cage multigenerational studies resulted in significantly lower populations of adult mosquitoes in cages containing treated DDALOs after 4 wk. Successful laboratory studies show that the DDALO appears to be a promising tool that could be used for controlling wild vector populations of Ae. aegypti in combination with other mosquito control practices.

Spatio-Temporal Distribution of Vector-Host Contact (VHC) Ratios and Ecological Niche Modeling of the West Nile Virus Mosquito Vector, Culex quinquefasciatus, in the City of New Orleans, LA, USA.

The consistent sporadic transmission of West Nile Virus (WNV) in the city of New Orleans justifies the need for distribution risk maps highlighting human risk of mosquito bites. We modeled the influence of biophysical and socioeconomic metrics on the spatio-temporal distributions of presence/vector-host contact (VHC) ratios of WNV vector, Culex quinquefasciatus, within their flight range. Biophysical and socioeconomic data were extracted within 5-km buffer radii around sampling localities of gravid female Culex quinquefasciatus. The spatio-temporal correlations between VHC data and 33 variables, including climate, land use-land cover (LULC), socioeconomic, and land surface terrain were analyzed using stepwise linear regression models (RM). Using MaxEnt, we developed a distribution model using the correlated predicting variables. Only 12 factors showed significant correlations with spatial distribution of VHC ratios (R² = 81.62, p < 0.01). Non-forested wetland (NFWL), tree density (TD) and residential-urban (RU) settings demonstrated the strongest relationship. The VHC ratios showed monthly environmental resilience in terms of number and type of influential factors. The highest prediction power of RU and other urban and built up land (OUBL), was demonstrated during May-August. This association was positively correlated with the onset of the mosquito WNV infection rate during June. These findings were confirmed by the Jackknife analysis in MaxEnt and independently collected field validation points. The spatial and temporal correlations of VHC ratios and their response to the predicting variables are discussed.

Field evaluation of a semi-automatic funnel trap targeted the medically important non-biting flies.

Bait-trapping is a useful approach for monitoring fly population dynamics, and it is an effective tool for physical control of pest species. The aim of this study was to test a newly developed semi-automatic funnel fly trap with some modifications of the former prototype fly trap to study medically important fly population density. The efficacy of the semi-automatic funnel trap was assessed by field sampling during July 2013-June 2014 using 1-day tainted beef offal as bait. The modified semi-automatic funnel traps were able to capture a total of 151,141 adult flies, belonging to the families: Calliphoridae (n=147,248; 97.4%), Muscidae (n=3,124; 2.1%) and Sarcophagidae (n=769; 0.5%), which are the medically important fly species. Among the total of 35 species collected, Chrysomya megacephala (Diptera: Calliphoridae) (n=88,273; 59.95%), Musca domestica (Diptera: Muscidae) (n=1,324; 42.38%) and Boettcherisca peregrina (Diptera: Sarcophagidae) (n=68; 33.01%) were the predominant species of each family. High number of flies was captured in forest area, representing 42.47% (n=64,197) of total specimens. Female flies were trapped more than male with total sex ratio of 0.37 male/female. Flies were trapped throughout the year with peak population in summer. Peak activity was recorded in the afternoon (12.00-18.00h). In summary, the modified semi-automatic funnel fly trap can be used for field collection of the adult fly. By setting the timer, population dynamics, diversity, and periodic activity of adult flies were determined.

Attractive Toxic Sugar Bait (ATSB) For Control of Mosquitoes and Its Impact on Non-Target Organisms: A Review.

Mosquito abatement programs contend with mosquito-borne diseases, insecticidal resistance, and environmental impacts to non-target organisms. However, chemical resources are limited to a few chemical classes with similar modes of action, which has led to insecticide resistance in mosquito populations. To develop a new tool for mosquito abatement programs that control mosquitoes while combating the issues of insecticidal resistance, and has low impacts of non-target organisms, novel methods of mosquito control, such as attractive toxic sugar baits (ATSBs), are being developed. Whereas insect baiting to dissuade a behavior, or induce mortality, is not a novel concept, as it was first introduced in writings from 77 AD, mosquito baiting through toxic sugar baits (TSBs) had been quickly developing over the last 60 years. This review addresses the current body of research of ATSB by providing an overview of active ingredients (toxins) include in TSBs, attractants combined in ATSB, lethal effects on mosquito adults and larvae, impact on non-target insects, and prospects for the use of ATSB.

Evaluations of dual attractant toxic sugar baits for surveillance and control of Aedes aegypti and Aedes albopictus in Florida.

BACKGROUND: Dual attractant toxic sugar baits (D-ATSB) containing two host kairomones, L-lactic (LA) and 1-octen-3-ol (O), and fruit-based attractants were evaluated through olfactory, consumption and mortality, and semi-field experiments to determine if host kairomones could first, enhance attraction of a fruit-based (attractant) toxic sugar bait (ATSB), and second, increase the efficacy of a fruit based attractive toxic sugar bait (ATSB). METHODS: Four combinations of LA and O were incorporated into the ATSB and evaluated in an olfactometer to determine if these combinations could enhance attraction of Aedes aegypti (L.) to the bait. Ae. albopictus (Skuse) and Ae. aegypti were used to determine bait consumption through excrement droplet counts and percent mortality, of the most attractive D-ATSB (1% LA and 1% O) from the olfactory study. Semi-field evaluations were conducted in screened portable field cages to determine if the D-ATSB applied to non-flowering plants controlled more mosquitoes than the fruit-based ATSB, and ASB. Mosquitoes were exposed to D-ATSB and the two controls for 48 h and collected with BGS traps. The catch rates of the BGS traps were compared to determine efficacy of the D-ATSB. RESULTS: During olfactometer evaluations of D-ATSB, Ae. aegypti mosquitoes were more attracted to 1% LA and 1% O compared to the fruit-based toxic sugar bait alone. Both species of mosquito consumed more fruit-based non-toxic bait (ASB) and ATSB than the D-ATSB. For both species, percent mortality bioassays indicated D-ATSB controlled mosquitoes, as compared to non-toxic control, but not more than the fruit based ATSB. Semi-field evaluations, BioGents sentinel traps at 48 h confirmed that ATSB (positive control) controlled Ae. albopictus, but there was no statistical difference between ASB (negative control) and the D-ATSB. No differences were observed between the mosquitoes caught in any of the experimental formulations for Ae. aegypti. CONCLUSIONS: L-lactic (1%) and 1-octen-3-ol (1%) added to a fruit-based sugar bait increased attraction of Ae. aegypti and may have future implications in mosquito trapping devices. The addition of the host kairomones did not enhance the consumption and efficacy of the ATSB in laboratory or semi-field evaluations for both mosquito species. We attribute to the absence of other host cues leading to lack of alighting onto bait surfaces to imbibe the toxic bait, as well as a possible decrease in palatability of the bait caused by the addition of the host kairomones.

Behavioral Responses of Nymph and Adult Cimex lectularius (Hemiptera: Cimicidae) to Colored Harborages.

Behavioral bioassays were conducted to determine whether bed bug adults and nymphs prefer specific colored harborages. Two-choice and seven-choice behavioral color assays indicate that red (28.5%) and black (23.4%) harborages are optimal harborage choices for bed bugs. Yellow and green harborages appear to repel bed bugs. Harborage color preferences change according to gender, nutritional status, aggregation, and life stage. Female bed bugs prefer harborages with shorter wavelengths (lilac-14.5% and violet-11.5%) compared to males, whereas males prefer harborages with longer wavelengths (red-37.5% and black-32%) compared with females. The preference for orange and violet harborages is stronger when bed bugs are fed as opposed to when they are starved. Lone bed bugs (30%) prefer to be in black harborages while red harborages appear to be the optimum harborage color for bed bugs in more natural mixed aggregations (35.5%). Bed bug nymphs preferred different colored harborages at each stage of development, which is indicative of their developing eye structures and pigments. First instars showed no significant preference for any colored harborage soon after hatching. However, by the fifth instar, 27.5% of nymphs significantly preferred red and black harborages (which was a similar preference to adult bed bugs). The proportion of oviposited eggs was significantly greater under blue, red, and black harborages compared to other colored harborages tested. The use of visual cues such as specific colors offers great potential for improving bed bug monitoring tools by increasing trap captures.

The Influence of Roughness and Pyrethroid Formulations on Bed Bug (Cimex lectularius L.) Resting Preferences.

Two-choice tests were conducted to examine the effect of surface roughness on the resting preference of bed bugs, Cimex lectularius L., on copper, basswood, and acrylic materials. The influence of pyrethroid formulation applications on resting preferences was also evaluated. Bed bugs were given the choice of resting between two sanded halves of each material tested. One half was sanded with a P60 grit sandpaper and the other with a less rough P600 grit sandpaper. A significantly higher proportion of bed bugs chose to rest on the rougher P60 grit sanded half of all materials tested. Pyrethroid applications were made to either the P60 grit half or both halves of acrylic arenas and resting preferences were again assessed. Behavioral responses of bed bugs to pyrethroid formulation applications varied depending on the bed bug strain used and the formulation applied. Bed bugs would still rest on the P60 grit half when Suspend SC formulation (0.06% deltamethrin) was applied; however, an avoidance response was observed from a bed bug strain susceptible to D-Force aerosol formulations (0.06% deltamethrin). The avoidance behavior is likely attributed to one, more than one, or even an interaction of multiple spray constituents and not the active ingredient.

Nontarget effects of aerial mosquito adulticiding with water-based unsynergized pyrethroids on honey bees and other beneficial insects in an agricultural ecosystem of north Greece.

We assessed the nontarget effects of ultra-low-volume (ULV) aerial adulticiding with two new water-based, unsynergized pyrethroid formulations, Aqua-K-Othrine (FFAST antievaporant technology, 2% deltamethrin) and Pesguard S102 (10% d-phenothrin). A helicopter with GPS navigation technology was used. One application rate was tested per formulation that corresponded to 1.00 g (AI)/ha of deltamethrin and 7.50 g (AI)/ha of d-phenothrin. Three beneficial nontarget organisms were used: honey bees (domesticated hives), family Apidae (Apis mellifera L.); mealybug destroyers, family Coccinellidae (Cryptolaemus montrouzieri Mulsant); and green lacewings, family Chrysopidae (Chrysoperla carnea (Stephens)). No significant nontarget mortalities were observed. No bees exhibited signs of sublethal exposure to insecticides. Beehives exposed to the insecticidal applications remained healthy and productive, performed as well as the control hives and increased in weight (25-30%), in adult bee population (14-18%), and in brood population (15-19%).

Droplet sampling of an oil-based and two water-based antievaporant ultra-low volume insecticide formulations using Teflon- and magnesium oxide-coated slides.

We estimated the diameters below which 50% and 90% of the volume of droplets exist (Dv50 and Dv90, respectively) of 1 oil-based (Permanone 30-30) and 2 water-based (AquaReslin, Aqua-K-Othrine) antievaporant aerosols (with the Film Forming Aqueous Spray Technology [FFAST]) using Teflon- and magnesium oxide (MgO)-coated slides and determined whether the aging of the droplets on the slides (up to 60 min) exhibited any significant effect on Dv50 and Dv90 calculations. There were no significant differences in either Dv50 or Dv90 estimates on MgO-coated slides at 0 min and 60 min for all 3 products tested. On Teflon-coated slides, the only product that showed significant difference between 0 min and 60 min in both Dv50 and Dv90 estimates was Aqua-K-Othrine, perhaps due to a difference in formulation components. Specifically, both values corresponding to Dv50 and Dv90 at 60 min decreased by approximately 50% when compared to the values at 0 min. For the other 2 products, AquaReslin and Permanone, aging of droplets on Teflon up to 60 min did not have any significant effect on Dv50 and Dv90 values. To further investigate the behavior of Aqua-K-Othrine droplets on Teflon-coated slides we observed the droplets immediately after spraying and at 10-min intervals under different conditions of temperature and humidity. The majority of the shrinkage occurred within the 1st 10 min after impaction on the slides under all conditions tested. So in most field situations where slides are read several hours or days after collection, this shrinkage would not be observed. The MgO-coated slides should be the preferred field method for sampling droplets of Aqua-K-Othirne with the FFAST antievaporant technology.

Efficacy of aerial ultra-low volume applications of two novel water-based formulations of unsynergized pyrethroids against riceland mosquitoes in Greece.

We assessed the efficacy of ultra-low volume aerial adulticiding with 2 new water-based, unsynergized formulations of Aqua-K-Othrin (2% deltamethrin) and Pesguard S102 (10% d-phenothrin) against the riceland mosquitoes of Greece. A helicopter with Global Positioning System (GPS) navigation, real-time weather recording, and spray dispersal modeling (AgDISP) was utilized to accurately treat the experimental blocks by adjusting spray line positions to changing meteorological conditions. Two application rates were applied per formulation that corresponded to 0.75 and 1.00 g AI/ha of deltamethrin and 7.50 and 10.00 g AI/ha of d-phenothrin. The mosquitoes used for the trials were the main nuisance species found in rice field areas of Thessaloniki, which were primarily Aedes caspius, followed by Culex modestus and Anopheles sacharovi. Overall mean mortality of caged mosquitoes was 69.2% and 64.8% for deltamethrin and d-phenothrin, respectively. Mean population decrease in wild mosquito populations within the treatment areas was 76.5% and 78% for deltamethrin and d-phenothrin, respectively. The AgDISP dispersal model, coupled with GPS navigation and real-time weather recording, enabled accurate placement of the spray cloud such that the majority of the treatment area received sufficiently high droplet densities to result in uniform caged-mosquito mortality across all sampling sites.

Vapor toxicity of three prototype volatile insecticidal compounds to house fly (Diptera: Muscidae).

Vapor toxicities of two prototype formate esters, ethylene glycol di-formate (EGDF) and heptyl formate, and the prototype heterobicyclic menthofuran were determined against laboratory-reared adult house flies, Musca domestica L. (Diptera: Muscidae). Toxicities were compared between house flies and other dipteran species tested previously. The organophosphate fumigant insecticide dichlorvos (DDVP) was used as a positive control. Although less toxic than DDVP, all three prototype compounds were toxic to house flies. Most toxic was menthofuran (LC50 = 3.70 mg/liter air), followed by EGDF (LC50 = 9.27 mg/liter air) and heptyl formate (LC50 = 32.62 mg/liter air). The toxicity of menthofuran when applied after piperonyl butoxide increased 1.5-fold. The toxicities of heptyl formate and EGDF when applied after S,S,S-tributyl phosphorotrithioate decreased by 1.5- and 2-folds, respectively. These results are slightly different from results obtained for other dipterans, further supporting previous hypotheses that toxicity may have some species specificity. Ceramic porous rods impregnated with heptyl formate were used to evaluate the effectiveness of controlled vapor release for a single dose of heptyl formate against house flies. Controlled vapor release ofheptyl formate can be used successfully to provide prolonged house fly mortality. Even though the prototype compounds did not exhibit toxicity as high as that of DDVP, their toxicities to dipterans, along with their reduced mammalian toxicities relative to DDVP, make them potential DDVP replacement candidates.

Effect of population structure and size on aggregation behavior of Cimex lectularius (Hemiptera: Cimicidae).

The bed bug, Cimex lectularius L. (Hemiptera: Cimicidae), occurs in aggregations until the conditions are no longer beneficial, leading to dispersal. Active and passive bed bug dispersal causes migrations from main aggregations either within a room, from room to room within a building, or from building to building. Because bed bug movement is an important factor in the spread of infestations, we wanted to determine how population structure and size affect bed bug aggregations. Engorged bed bugs were placed in glass petri dish arenas at varying densities, sex ratios, and population compositions. Nymphs had a high tendency to aggregate, varying between 94 and 98%, and therefore were not the likely dispersal stage of the bed bug. At densities of 10 and 40 adults at a 1:1 sex ratio, there were significantly more lone females than lone males. When the population composition was varied, the percentage of lone females was significantly higher than that of males and nymphs at population compositions of 40 and 80% adults. When the sex ratio of adults was varied, there were significantly more lone females than males in arenas with 20, 50, and 80% males. Females, being found away from aggregations significantly more often than any other life stage, are potentially the dispersal stage of the bed bug. Active female dispersal away from main aggregations can potentially lead to treatment failures and should be taken into account when using control methods.

Lethal effects of heat and use of localized heat treatment for control of bed bug infestations.

Bed bugs, Cimex lectularius L., hide in cracks and crevices in furniture and are difficult to control. The bed bug thermal death kinetics were examined to develop a heat treatment method to eliminate bed bug infestations in room contents. High temperatures caused temporary immobilization (knockdown) of bed bugs even with exposures that did not have lethal effects. Exposure of bed bug adults to 39 degrees C for 240 min caused no mortality; however, as temperatures increased from 41 to 49 degrees C, exposure times that caused 100% mortality decreased. The temperature difference to provide a 10-fold change in the mortality was estimated at 4 degrees C, and the estimated activation energy (EA) was between 484 and 488.3 kJ/mol. This demonstrates that bed bugs are not more resistant or susceptible to changes in temperature than other tested insects and that the temperatures needed to kill bed bugs are relatively low. In room treatment tests, heat treatment times varied from 2 to 7 h with complete mortality of exposed bed bugs within the treatment envelope created by surrounding the treated furniture with polystyrene sheathing boards. Containment and circulation of heat around the treated material were crucial factors in an efficient heat treatment for bed bug control. The room floor material greatly affected containment of the heat. The tested method for limited heat treatment of furniture and other room contents required equipment costing less than US$400 and provided opportunity for residual pesticide application around the room with minimal disruption in use of treated room.

Toxicities of 31 volatile low molecular weight compounds against Aedes aegypti and Culex quinquefasciatus.

This research studied 31 volatile compounds for indoor control of the medically important mosquitoes Aedes aegypti L. and Culex quinquefasciatus Say. Only adult female mosquitoes were tested. The test compounds were from six families that included five heterobicyclics, eight formate esters, formic acid (a hydrolyzed metabolite of formate esters), eight acetate esters, four propionate esters, three butyrate esters, and two valerate esters. Also, the organophosphate compound dichlorvos (DDVP) was tested as a positive control. Cx. quinquefasciatus was generally more susceptible than Ae. aegypti. Cx. quinquefasciatus was most susceptible to a subset of heterobicyclics and formate esters (rank: n-butyl formate > hexyl formate = dihydrobenzofuran = menthofuran = heptyl formate = ethyl formate). Ae. aegypti was most susceptible to a subset of formate esters (rank: methyl > n-butyl > propyl = ethyl = hexyl). The most active materials against both species had LC50s of 0.4-1 mg active ingredient per 0.5 liter of air volume (0.8-2 mg/liter), which is 50- to 60-fold less toxic than dichlorvos (an organophosphate insecticide that is being phased out from indoor use). In relation to Drosophila melanogaster Meigen, both mosquito species were generally more susceptible to formate esters but more tolerant of heterobicyclics. Generally, the most toxic compound against all dipterans tested to date is n-butyl formate, whereas menthofuran is additionally toxic against Cx. quinquefasciatus and D. melanogaster. Finally, the toxicity differences between species point to the potential for differential toxicity among mosquito general/species, suggesting that further studies of a number of mosquito species might be warranted.

Ability of bed bug-detecting canines to locate live bed bugs and viable bed bug eggs.

The bed bug, Cimex lectularius L., like other bed bug species, is difficult to visually locate because it is cryptic. Detector dogs are useful for locating bed bugs because they use olfaction rather than vision. Dogs were trained to detect the bed bug (as few as one adult male or female) and viable bed bug eggs (five, collected 5-6 d after feeding) by using a modified food and verbal reward system. Their efficacy was tested with bed bugs and viable bed bug eggs placed in vented polyvinyl chloride containers. Dogs were able to discriminate bed bugs from Camponotus floridanus Buckley, Blattella germanica (L.), and Reticulitermes flavipes (Kollar), with a 97.5% positive indication rate (correct indication of bed bugs when present) and 0% false positives (incorrect indication of bed bugs when not present). Dogs also were able to discriminate live bed bugs and viable bed bug eggs from dead bed bugs, cast skins, and feces, with a 95% positive indication rate and a 3% false positive rate on bed bug feces. In a controlled experiment in hotel rooms, dogs were 98% accurate in locating live bed bugs. A pseudoscent prepared from pentane extraction of bed bugs was recognized by trained dogs as bed bug scent (100% indication). The pseudoscent could be used to facilitate detector dog training and quality assurance programs. If trained properly, dogs can be used effectively to locate live bed bugs and viable bed bug eggs.