Determination of the Discriminating Concentration Towards Permethrin for Surveying Resistance in Amblyomma americanum.

Amblyomma americanum Linnaeus (Ixodida: Ixodidae) is ubiquitously present throughout the southeastern United States and is capable of vectoring several pathogens. White-tailed deer are the main host for adult A. americanum. However, this tick species is a generalist that will feed on most vertebrates, including humans, deer, livestock, and pets. Management of this species can be challenging due to a lack of cost-effective strategies. Acaricides are often utilized, however, this may lead to pesticide resistance. The Food and Agriculture Organization of the United Nations (FAO) larval packet test (LPT) was performed on susceptible A. americanum to determine the lethal concentration (LC) and discriminating concentration (DC) values for permethrin. The FAO LPT was used at these pre-established values to compare levels of resistance in ticks collected from a captive deer farm and wild areas representing high and low permethrin exposure settings, respectively. Resistance ratios (RR) calculated from the LC values for the ticks collected from farmed and wild deer ranged between 1 and 2. A. americanum collected from farmed and wild deer were not found to be resistant, however, some samples had slightly elevated RRs as compared to the susceptible laboratory strain, which may suggest tolerance development. Although the A. americanum sampled in this study were not resistant to permethrin, the DC calculated in this study will allow for rapid evaluation of resistance in a permethrin resistance monitoring program such that alternate management strategies can be adopted if resistance is detected.

Prevalence and distribution of pathogen infection and permethrin resistance in tropical and temperate populations of Rhipicephalus sanguineus s.l. collected worldwide.

The brown dog tick, Rhipicephalus sanguineus sensu lato (s.l.) Latreille (Acari: Ixodidae), is a peridomestic, cosmopolitan parasite of dogs known to vector numerous pathogens of veterinary and medical importance. Recent phylogenetic analyses separate this tick into temperate and tropical lineages. Populations of Rh. sanguineus s.l. have been reported to exhibit sodium channel target site insensitivity to permethrin and etofenprox, which is likely due to the prolonged use of pyrethroids against many pests in and around the home. In this study, populations collected in the Caribbean, Africa, Asia, Europe and North America, were tested to identify the distribution of a known resistance mechanism, pathogen-vector interactions and phylogeny in relation to latitude. Using molecular assays, populations from 29 distinct locations were simultaneously geographically typed and screened for bacterial infection by Rickettsia, Ehrlichia, Babesia and Hepatozoon species, and for the presence of a sodium channel single nucleotide polymorphism known to confer permethrin resistance. Implications of these results on Rh. sanguineus s.l. management in association with geographical distribution will be discussed.

Electrophysiologically and behaviourally active semiochemicals identified from bed bug refuge substrate.

Bed bugs are pests of public health importance due to their relentless biting habits that can lead to allergies, secondary infections and mental health issues. When not feeding on human blood bed bugs aggregate in refuges close to human hosts. This aggregation behaviour could be exploited to lure bed bugs into traps for surveillance, treatment efficacy monitoring and mass trapping efforts, if the responsible cues are identified. The aim of this study was to identify and quantify the bed bug aggregation pheromone. Volatile chemicals were collected from bed bug-exposed papers, which are known to induce aggregation behaviour, by air entrainment. This extract was tested for behavioural and electrophysiological activity using a still-air olfactometer and electroantennography, respectively. Coupled gas chromatography-electroantennography (GC-EAG) was used to screen the extract and the GC-EAG-active chemicals, benzaldehyde, hexanal, (E)-2-octenal, octanal, nonanal, decanal, heptanal, (R,S)-1-octen-3-ol, 3-carene, ß-phellandrene, (3E,5E)-octadien-2-one, (E)-2-nonenal, 2-decanone, dodecane, nonanoic acid, 2-(2-butoxyethoxy)ethyl acetate, (E)-2-undecanal and (S)-germacrene D, were identified by GC-mass spectrometry and quantified by GC. Synthetic blends, comprising 6, 16, and 18 compounds, at natural ratios, were then tested in the still-air olfactometer to determine behavioural activity. These aggregation chemicals can be manufactured into a lure that could be used to improve bed bug management.

Auto-dissemination of commercially available fungal pathogens in a laboratory assay for management of the brown dog tick Rhipicephalus sanguineus.

Rhipicephalus sanguineus is a canine tick that infests dogs throughout the world and is frequently found in homes and dog kennels. Management of this tick species is complicated by the presence of resistance to commonly utilized acaricides. Fungal formulations could provide a valuable alternative tool for management and are especially relevant indoors where detrimental environmental effects on fungal spores are of less concern. Two commercially available fungal formulations, one containing Metarhizium anisopliae and the other containing Beauveria bassiana, were compared for time to death and sporulation in nymphal ticks exposed for 60 min in treated filter paper packets. Beauveria bassiana exposure killed ticks faster than M. anisopliae exposure and B. bassiana was more likely to sporulate on tick cadavers than M. anisopliae. To determine whether infected ticks could disseminate fungus to their conspecifics, ticks were marked and treated with fungus before being placed with untreated ticks. Fungus was successfully transmitted from treated to untreated ticks. Mortality of ticks exposed to B. bassiana-exposed conspecifics occurred sooner than for those exposed to M. anisopliae-exposed conspecifics, indicating faster dissemination in the former. Therefore, although both formulations resulted in decreased longevity of ticks compared with the controls, the B. bassiana formulation holds the most promise for direct or indirect application with respect to brown dog tick management.

Effects of four commercial fungal formulations on mortality and sporulation in house flies (Musca domestica) and stable flies (Stomoxys calcitrans).

The house fly Musca domestica L. (Diptera: Muscidae) and stable fly Stomoxys calcitrans (L.) (Diptera: Muscidae) are major pests of livestock. Biological control is an important tool in an integrated control framework. Increased mortality in filth flies has been documented with entomopathogenic fungi, several strains of which are commercially available. Three strains of Beauveria bassiana (Balsamo-Crivelli) Vuillemin (Hypocreales: Cordycipitaceae) and one strain of Metarhizium brunneum (Petch) (Hypocreales: Clavicipitaceae) were tested in commercial formulations for pathogenicity against house flies and stable flies. There was a significant increase in mortality of house flies with three of the formulations, BotaniGard® ES, Mycotrol® O, and Met52® EC, during days 4-9 in comparison with balEnce™ and the control. In stable flies, mortality rates were highest with Met52® EC, followed by Mycotrol® O, BotaniGard® ES and, finally, balEnce™. There was a significant fungal effect on sporulation in both house flies and stable flies. Product formulation, species differences and fungal strains may be responsible for some of the differences observed. Future testing in field situations is necessary. These commercial biopesticides may represent important tools in integrated fly management programmes.

Vision in the common bed bug Cimex lectularius L. (Hemiptera: Cimicidae): eye morphology and spectral sensitivity.

Bed bugs as pests of public health importance recently experienced a resurgence in populations throughout the U.S. and other countries. Consequently, recent research efforts have focused on improving understanding of bed bug physiology and behaviour to improve management. While few studies have investigated the visual capabilities of bed bugs, the present study focused specifically on eye morphology and spectral sensitivity. A 3-D imaging technique was used to document bed bug eye morphology from the first instar through adult and revealed morphological characteristics that differentiate the common bed bug from the tropical bed bug as well as sex-specific differences. Electrophysiological measurements were used to evaluate the spectral sensitivity of adult bed bugs. Male bed bugs were more responsive than females at some wavelengths. Electrophysiological studies provided evidence for at least one photoreceptor with a spectral sensitivity curve peak in the green (λmax 520 nm) region of the spectrum. The broadened long wavelength portion of the spectral sensitivity curve may potentially indicate another photoreceptor in the yellow-green (λmax 550 nm) portion of the spectrum or screening pigments. Understanding more about bed bug visual biology is vital for designing traps, which are an important component of integrated bed bug management.

Oviposition Deterrence and Immature Survival of Filth Flies (Diptera: Muscidae) When Exposed to Commercial Fungal Products.

Filth flies are pests of livestock, and can transmit pathogens that cause disease to animals and their caretakers. Studies have shown successful infection of adult filth flies following exposure to different strains and formulations of entomopathogenic fungi. This study aimed to examine the effects of commercial formulations of Beauveria bassiana (Balsamo) (Moniliales: Moniliaceae) (i.e., BotaniGard ES, Mycotrol O, balEnce), and Metarhizium brunneum (Metsch.) (Ascomycota: Hypocreales) (i.e., Met52 EC), on filth fly oviposition and immature fly survival after exposure. House flies, Musca domestica L., laid significantly fewer eggs on Met52 EC-treated surfaces than on surfaces treated with all other products and the control. Similar numbers of eggs were laid on surfaces treated with all B. bassiana products, but egg production was half of the control. Stable flies, Stomoxys calcitrans (L.), laid the fewest eggs on Met52 EC- and Mycotrol O-treated surfaces. This species did not distinguish between the remaining products and the control. In a second experiment, house fly eggs were placed on treated cloths so that hatched larvae contacted the treatment prior to development. Met52 EC had the greatest effect on immature survival with a significant reduction in recovered pupae at the medium and high doses of fungi. Overall, Met52 EC, containing M. brunneum, had the greatest effect on house fly and stable fly oviposition deterrence and immature development of house flies. Management implications are discussed.

Community-based sampling methods for surveillance of the Chagas disease vector, Triatoma dimidiata (Hemiptera: Reduviidae: Triatominae).

In Guatemala, the most widespread vector of Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae), the causative agent of Chagas disease, is Triatoma dimidiata (Latreille) (Hemiptera: Reduviidae: Triatominae). T. dimidiata is native to Guatemala and is present in both domestic and sylvatic habitats. Consequently, control of T. dimidiata is difficult because after successful elimination from homes, individual insects can recolonize homes from the surrounding environment. Therefore, intensive long-term surveillance of this species is essential to ensure adequate control is achieved. Manual inspection for signs of infestation, the current method used to monitor Triatominae throughout Central and South America, is labor and time-consuming, so cost-effective alternatives are needed. The current study compared the effectiveness of the current method of surveillance of T. dimidiata with community-based techniques of G6mez-Nuñez sensor boxes, collection and observation of bugs by householders, and presence of triatomine-like feces on walls. Although manual inspection was the most sensitive method when used alone, collection by householders also was sensitive and specific and involved less effort. Sensor boxes were not sensitive indicators of T. dimidiata infestation when used alone. Two recorded variables, visual inspection for feces and the sighting of bugs by householders, were sensitive and specific indicators of infestation, and in combination with collection by householders and sensor boxes these methods were significantly more likely to detect infestations than manual inspection alone. A surveillance program that combines multiple community-based techniques should have low cost and involve minimal effort from the government and at the same time promote sustainable community involvement in disease prevention.

Risk factors for domestic infestation by the Chagas disease vector, Triatoma dimidiata in Chiquimula, Guatemala.

In Guatemala prior to control initiatives, the main vectors of Trypanosoma cruzi, the causative agent of Chagas disease, were Rhodnius prolixus and Triatoma dimidiata. This study conducted in 2006 in the department of Chiquimula recorded a high level of T. dimidiata infestation and an absence of R. prolixus in all surveyed communities. In Guatemala, the presence of T. dimidiata as domestic, peridomestic and sylvatic populations results in control difficulties as houses are re-infested from the surrounding environment. Entomological surveys, the current method used to select houses in need of control efforts, are labour intensive and time consuming. A time- and cost-effective way to prioritize houses for evaluation and subsequent treatment is the stratification of houses based on the risk of triatomine infestation. In the present study, 17 anthropogenic risk factors were evaluated for associations with house infestation of T. dimidiata including: wall, floor and roof type. There was an increased likelihood of domestic infestation with T. dimidiata associated with the presence of dirt floors (18/29; OR 8.075, 95% CI 2.13-30.6), uncoated bajareque walls (12/17; OR 4.80, 95% CI 1.35-17.1) and triatomine-like faeces on walls (16/26; OR 3.89, 95% CI 1.19-12.7). These factors could be used to target control of T. dimidiata to communities with an increased risk of being infested.

Tracking bed bugs (Cimex lectularius): a study of the effect of physiological and extrinsic factors on the response to bed bug-derived volatiles.

The common bed bug, Cimex lectularius, feeds on the blood of mammal and bird hosts, and is a pest of global importance. Semiochemicals are chemicals involved in animal communication that may affect behaviour and/or physiology. Attractive semiochemicals that play a role in mediating bed bug behaviour could be exploited for the development of a highly effective novel monitoring device. Tracking software was used to record the response of bed bugs to volatiles from paper previously exposed to conspecific bugs in a still-air olfactometer illuminated by infrared lights, through a variety of activity variables. The effect of time of day as an extrinsic factor, and sex, stage, mating status and nutritional status as physiological factors on the response of bed bugs to the volatiles was examined. Bed bugs of both sexes and all stages responded to the volatiles from bed bug-exposed papers, showing significant attraction and orientation towards the volatile source whether they were starved or engorged. Confirmation that the physiological factors examined do not affect the response of bed bugs to the volatiles from bed bug-exposed papers provides evidence that these bed bug-derived volatiles contain aggregation cues, as semiochemicals that promote aggregation should by definition be detected by both sexes and all life stages. A device baited with such semiochemicals could play a major role in limiting the impact of the current bed bug resurgence by enabling timely detection of infestations, along with quantitative evaluation of control and effective surveillance of the geographical distribution of the pest species.

A bioassay for studying behavioural responses of the common bed bug, Cimex lectularius (Hemiptera: Cimicidae) to bed bug-derived volatiles.

The common bed bug, Cimex lectularius (Hemiptera: Cimicidae), has recently re-emerged in increasing numbers, distribution and intensity of infestation in many countries. Current control relies on the application of residual pesticides; but, due to the development of insecticide resistance, there is a need for new tools and techniques. Semiochemicals (behaviour and physiology modifying chemicals) could be exploited for management of bed bugs. However, in order to identify semiochemicals that can be utilised in monitoring or control, a suitable olfactometer is needed that enables the study of the responses of bed bugs to volatile chemicals. Previous studies have used olfactometers that do not separate olfactory responses from responses to physical contact. In this study, a still-air olfactometer was used to measure behavioural responses to different bed bug-derived volatiles presented in an odour pot. Bed bugs were significantly more likely to visit the area above the odour pot first, and more frequently, in the presence of volatiles from bed bug-exposed paper but not in the presence of volatiles from conspecific bed bugs. Bed bug activity was found to be dependent on the presence of the volatiles from bed bug-exposed paper, the time during the scotophase and the sex of the insect being tested. The still-air olfactometer could be used to test putative semiochemicals, which would allow an understanding of their behavioural role in bed bug ecology. Ultimately, this could lead to the identification of new semiochemical tools for bed bug monitoring and control.