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.

Species-specific alterations in Anopheles mosquito olfactory responses caused by Plasmodium infection.

Mosquitoes infected with malaria parasites have demonstrated altered behaviour that may increase the probability of parasite transmission. Here, we examine the responses of the olfactory system in Plasmodium falciparum infected Anopheles gambiae, Plasmodium berghei infected Anopheles stephensi, and P. berghei infected An. gambiae. Infected and uninfected mosquitoes showed differential responses to compounds in human odour using electroantennography coupled with gas chromatography (GC-EAG), with 16 peaks triggering responses only in malaria-infected mosquitoes (at oocyst, sporozoite or both stages). A selection of key compounds were examined with EAG, and responses showed differences in the detection thresholds of infected and uninfected mosquitoes to compounds including lactic acid, tetradecanoic acid and benzothiazole, suggesting that the changes in sensitivity may be the reason for differential attraction and biting at the oocyst and sporozoite stages. Importantly, the different cross-species comparisons showed varying sensitivities to compounds, with P. falciparum infected An. gambiae differing from P. berghei infected An. stephensi, and P. berghei infected An. gambiae more similar to the P. berghei infected An. stephensi. These differences in sensitivity may reflect long-standing evolutionary relationships between specific Plasmodium and Anopheles species combinations. This highlights the importance of examining different species interactions in depth to fully understand the impact of malaria infection on mosquito olfactory behaviour.

Mosquitoes (Diptera: Culicidae) Collected From Residential Yards and Dog Kennels in Florida Using Two Aspirators, a Sweep Net, or a CDC Trap.

Mosquito surveillance typically uses Centers for Disease Control and Prevention (CDC) mosquito light traps baited with CO2. From January 2013 to March 2015, we sampled seven field sites using three active mosquito-trapping techniques (two different aspirators and a sweep net) and the stationary CO2-baited CDC mosquito light trap to determine mosquito capture efficacy for each technique. Sampling occurred in four suburban backyards and three dog kennel facilities near Gainesville, FL, USA; species collection and relative abundance were measured. A total of 32 species and 70,090 individual mosquitoes were collected, including a new record for Alachua County, Florida, Aedes hendersoni (Cockerell). The dominant (>5% of total capture) mosquito species collected during the study included Aedes atlanticus (Dyar and Knab), Aedes infirmatus (Dyar and Knab), Anopheles crucians Wiedemann, Culiseta melanura (Coquillett), Culex erraticus (Dyar and Knab), Culex nigripalpus Theobald, and Uranotaenia sapphirina (Osten Sacken). The CDC trap captured the most species (29), followed by large aspirator (28), small aspirator (26), and the sweep net (23). All dominant species were captured with each sampling technique. Excluding Wyeomyia mitchellii (Theobald), all subdominant species (1-5% of total capture) were collected with each sampling technique. Future sampling should consider the utility (e.g., large numbers are readily collected) and limitations (e.g., personnel requirements) of aspirator collections when designing field-based mosquito sampling projects, especially those in residential areas or those focused upon species captured.

Next generation crop models: A modular approach to model early vegetative and reproductive development of the common bean (Phaseolus vulgaris L).

The next generation of gene-based crop models offers the potential of predicting crop vegetative and reproductive development based on genotype and weather data as inputs. Here, we illustrate an approach for developing a dynamic modular gene-based model to simulate changes in main stem node numbers, time to first anthesis, and final node number on the main stem of common bean (Phaseolus vulgaris L.). In the modules, these crop characteristics are functions of relevant genes (quantitative trait loci (QTL)), the environment (E), and QTL × E interactions. The model was based on data from 187 recombinant inbred (RI) genotypes and the two parents grown at five sites (Citra, FL; Palmira, Colombia; Popayan, Colombia; Isabela Puerto Rico; and Prosper, North Dakota). The model consists of three dynamic QTL effect models for node addition rate (NAR, No. d- 1), daily rate of progress from emergence toward flowering (RF), and daily maximum main stem node number (MSNODmax), that were integrated to simulate main stem node number vs. time, and date of first flower using daily time steps. Model evaluation with genotypes not used in model development showed reliable predictions across all sites for time to first anthesis (R2 = 0.75) and main stem node numbers during the linear phase of node addition (R2 = 0.93), while prediction of the final main stem node number was less reliable (R2 = 0.27). The use of mixed-effects models to analyze multi-environment data from a wide range of genotypes holds considerable promise for assisting development of dynamic QTL effect models capable of simulating vegetative and reproductive development.

Permethrin and malathion LD90 values for Culex quinquefasciatus vary with topical application site.

Prior research in multiple insect species has demonstrated that insecticide-induced mortality varies according to the body region exposed on the insect. This variation has been demonstrated in Culex quinquefasciatus Say (Diptera: Culicidae), but has not been quantified using dose-response curves. Applications of technical permethrin or malathion to one of three body regions on Cx. quinquefasciatus resulted in dose-response curves that were not equivalent to one another. The generated LD90 values and curves for each body region were compared with previously reported LD values for analogous sites in several mosquito species, specifically the mesothorax. Based on the present results, the permethrin and malathion LD50 and LD90 concentrations required for droplets impinging on the abdomen and mesothorax of Cx. quinquefasciatus when applied through ground-based spray systems utilized by mosquito control programmes were calculated.

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.

Climbing Ability of the Common Bed Bug (Hemiptera: Cimicidae).

Little is known about what factors influence the climbing ability of bed bugs, Cimex lectularius L. (Hemiptera: Cimicidae), in relation to the various surfaces they encounter. We examined how sex, time since last fed, and what surfaces the bed bugs were in contact with affected their climbing performance. The effects of sex and time since fed were tested by counting the number of bed bugs able to climb a 45° slope. The pulling force was recorded using an analytical balance technique that captured the sequential vertical pulling force output of bed bugs attached to various surfaces. Recently fed female bed bugs were found to have the most difficulty in climbing smooth surfaces in comparison with males. This difference can be explained by the larger weight gained from bloodmeals by female bed bugs. A variety of vertical pulling forces were observed on surfaces ranging from sandpaper to talc powder-covered glass. For surfaces not treated with talc powder, bed bugs generated the least amount of vertical pulling force from synthetically created 0.6-µm plastron surfaces. This vast range in the ability of bed bugs to grip onto various surfaces may have implications on limiting bed bugs dispersal and hitchhiking behaviors.

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.

Combining cattle and wound-derived synthetic attractants, POC and Bezzilure B, for sampling Chrysomya bezziana in Indonesia.

The myiasis fly Chrysomya bezziana Villeneuve (Diptera: Calliphoridae) is an obligate parasite of body orifices and unnatural openings or wounds of vertebrates. It is a major pest of livestock and responsible for economic losses throughout the Old World. Two chemical lures were tested in the present study to assess their effectiveness at trapping Ch. bezziana. Bezzilure B contains a synthetic lure of 2-mercaptoethanol originally identified from cattle wound volatiles. POC is a synthetic lure comprising 4-methyl phenol, 3-propyl phenol and racemic 1-octen-3-ol, originally identified from volatiles of ox odour and now used to control tsetse flies (Diptera: Glossina). Bezzilure B and POC lures were tested alone, and in combination, in a laboratory free flight room bioassay and in a field trial. Sticky traps were used to capture males and females of Ch. bezziana and to determine the efficacy of the lures. In the laboratory, Bezzilure B caught significantly more male and female flies than Bezzilure B+POC (p<0.001). Similarly, in the field, treatment significantly affected the total number of Ch. bezziana caught (p<0.001) but, contrastingly, Bezzilure B+POC caught significantly more flies than Bezzilure B (p<0.05). The differing results between the laboratory and field may simply reflect the different assays and could suggest different roles of semiochemicals at different operational distances. Although ineffective on its own, POC has a synergistic effect with Bezzilure B to increase fly catches in the field: by proportion, Bezzilure B+POC was 2.17 times as efficient as Bezzilure B for Ch. bezziana, and 1.16-2.68 times as efficient for other flies depending on the species collected.

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.

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.