A novel prolixicin identified in common bed bugs with activity against both bacteria and parasites.

The hematophagous common bed bug, Cimex lectularius, is not known to transmit human pathogens outside laboratory settings, having evolved various immune defense mechanisms including the expression of antimicrobial peptides (AMPs). We unveil three novel prolixicin AMPs in bed bugs, exhibiting strong homology to the prolixicin of kissing bugs, Rhodnius prolixus, and to diptericin/attacin AMPs. We demonstrate for the first time sex-specific and immune mode-specific upregulation of these prolixicins in immune organs, the midgut and rest of body, following injection and ingestion of Gr+ (Bacillus subtilis) and Gr- (Escherichia coli) bacteria. Synthetic CL-prolixicin2 significantly inhibited growth of E. coli strains and killed or impeded Trypanosoma cruzi, the Chagas disease agent. Our findings suggest that prolixicins are regulated by both IMD and Toll immune pathways, supporting cross-talk and blurred functional differentiation between major immune pathways. The efficacy of CL-prolixicin2 against T. cruzi underscores the potential of AMPs in Chagas disease management.

Activation of immune pathways in common bed bugs, Cimex lectularius, in response to bacterial immune challenges - a transcriptomics analysis.

The common bed bug, Cimex lectularius, is an urban pest of global health significance, severely affecting the physical and mental health of humans. In contrast to most other blood-feeding arthropods, bed bugs are not major vectors of pathogens, but the underlying mechanisms for this phenomenon are largely unexplored. Here, we present the first transcriptomics study of bed bugs in response to immune challenges. To study transcriptional variations in bed bugs following ingestion of bacteria, we extracted and processed mRNA from body tissues of adult male bed bugs after ingestion of sterile blood or blood containing the Gram-positive (Gr+) bacterium Bacillus subtilis or the Gram-negative (Gr-) bacterium Escherichia coli. We analyzed mRNA from the bed bugs' midgut (the primary tissue involved in blood ingestion) and from the rest of their bodies (RoB; body minus head and midgut tissues). We show that the midgut exhibits a stronger immune response to ingestion of bacteria than the RoB, as indicated by the expression of genes encoding antimicrobial peptides (AMPs). Both the Toll and Imd signaling pathways, associated with immune responses, were highly activated by the ingestion of bacteria. Bacterial infection in bed bugs further provides evidence for metabolic reconfiguration and resource allocation in the bed bugs' midgut and RoB to promote production of AMPs. Our data suggest that infection with particular pathogens in bed bugs may be associated with altered metabolic pathways within the midgut and RoB that favors immune responses. We further show that multiple established cellular immune responses are preserved and are activated by the presence of specific pathogens. Our study provides a greater understanding of nuances in the immune responses of bed bugs towards pathogens that ultimately might contribute to novel bed bug control tactics.

Bimodal alarm signals modulate responses to monomodal alarm signals in Camponotus modoc carpenter ants.

Distressed western carpenter ants, Camponotus modoc, produce alarm pheromone and substrate-borne vibrations. The alarm pheromone attracts nestmates but the effects of vibratory signals, or of bimodal pheromonal and vibratory signals, are not known. Worker ants of two Camponotus congeners reportedly stand still ("freeze") or run fast in response to engineered drumming vibrations inputted on plastic, but many responses to ant-produced vibratory signals on wood have not yet been investigated. Generally, orientating toward signalers under vertebrate predator attack seems maladaptive and not beneficial to ant colonies. We tested the hypotheses (1) that vibratory alarm signals cause freezing, rapid running but not attraction of nestmates, and (2) that bimodal alarm signals modulate responses to monomodal alarm signals, thereby possibly reducing predation risk. Laser Doppler vibrometry recordings revealed that the ants' vibratory signals readily propagate through ant nest lamellae, and thus quickly inform nest mates of perceived threats. With a speaker modified to record and deliver vibratory signals, we obtained drumming signals of distressed ants on a Douglas fir veneer, and bioassayed signal effects on ants in an arena with a suspended veneer floor. In response playback of vibratory signals, ants ran rapidly, or froze, but did not approach the vibratory signals. Exposed to alarm pheromone, ants frequently visited the pheromone source. However, concurrently exposed to both alarm pheromone and vibratory signals, ants visited the pheromone source less often but spent more time "frozen." The ants' modulated responses to bimodal signals seem adaptive but the reproductive fitness benefits are still to be quantified.

Carbon Dioxide, Methane, and Synthetic Cattle Breath Volatiles Attract Host-Seeking Stable Flies, Stomoxys calcitrans.

Stable flies, Stomoxys calcitrans (L.), are blood-feeding ectoparasites of cattle. Host-seeking stable flies respond to various cattle host cues, but a potential role of cattle breath gases [carbon dioxide (CO2), methane (CH4)] and cattle breath volatiles (acetone, isoprene, 2-butanone, 2-propanol, propionic acid, 3-methyl butyric acid, phenol), alone or in combination, on host-seeking behavior of stable flies has not yet been comprehensively investigated. In laboratory and greenhouse experiments, we tested the hypotheses that (1) CO2 and CH4 interactively attract stable flies, (2) CO2 'gates' attraction of stable flies to CH4, and (3) breath volatiles on their own, or in combination with both CO2 and CH4, attract stable flies. In Y-tube olfactometer experiments, the blend of CH4 (0.5%) and CO2 (1%) in breathing air ('b-air') attracted significantly more female flies than CH4, or CO2, in b-air. The flies' responses to CH4 were contingent upon their prior or concurrent exposure to CO2. In two-choice experiments in a large greenhouse compartment, significantly more flies landed on the host-look-alike barrel that disseminated a blend of CO2 and CH4 in b-air (CO2/CH4/b-air) than on the barrel disseminating either b-air or CO2. Moreover, significantly more flies landed on the barrel that disseminated synthetic breath volatiles (SBVs) than on the barrel disseminating b-air. The blend of CO2/CH4/b-air and SBVs elicited more fly landings on barrels than CO2/CH4/b-air but not than SBVs. SBVs, possibly combined with both CH4 and CO2, could be developed as a lure to enhance trap captures of stable flies in livestock production facilities.

Black blow fly (Diptera: Calliphoridae) bacterial symbionts inform oviposition site selection by stable flies (Diptera: Muscidae).

Larval habitats of blood-feeding stable flies, Stomoxys calcitrans (L.) (Diptera: Muscidae), overlap with foraging sites of black blow flies, Phormia regina (Meigen) (Diptera: Calliphoridae). We tested the hypothesis that bacteria in blow fly excreta inform oviposition decisions by female stable flies. In laboratory 2-choice bioassays, we offered gravid female stable flies fabric-covered agar plates as oviposition sites that were kept sterile or inoculated with either a blend of 7 bacterial strains isolated from blow fly excreta (7-isolate-blend) or individual bacterial isolates from that blend. The 7-isolate-blend deterred oviposition by female stable flies, as did either of 2 strains of Morganella morganii subsp. sibonii. Conversely, Exiguobacterium sp. and Serratia marcescens each prompted oviposition by flies. The flies' oviposition decisions appear to be guided by bacteria-derived semiochemicals as the bacteria could not be physically accessed. Oviposition deterrence caused by semiochemicals of the 7-isolate-blend may help stable flies avoid competition with blow flies. The semiochemicals of bioactive bacterial strains could be developed as trap lures to attract and capture flies and deter their oviposition in select larval habitats.

Effects of macro- and micro-nutrients on momentary and season-long feeding responses by select species of ants.

Few studies have investigated the relative contribution of specific nutrients to momentary and season-long foraging responses by ants. Using western carpenter ants, Camponotus modoc, and European fire ants, Myrmica rubra, as model species, we: (1) tested preferential consumption of various macro- and micro-nutrients; (2) compared consumption of preferred macro-nutrients; (3) investigated seasonal shifts (late May to mid-September) in nutrient preferences; and (4) tested whether nutrient preferences of C. modoc and M. rubra pertain to black garden ants, Lasius niger, and thatching ants, Formica aserva. In laboratory and field experiments, we measured nutrient consumption by weighing Eppendorf tubes containing aqueous nutrient solutions before and after feeding by ants. Laboratory colonies of C. modoc favored nitrogenous urea and essential amino acids (EAAs), whereas M. rubra colonies favored sucrose. Field colonies of C. modoc and M. rubra preferentially consumed EAAs and sucrose, respectively, with no sustained shift in preferred macro-nutrient over the course of the foraging season. The presence of a less preferred macro-nutrient in a nutrient blend did not diminish the blend's 'appeal' to foraging ants. Sucrose and EAAs singly and in combination were equally consumed by L. niger, whereas F. aserva preferred EAAs. Baits containing both sucrose and EAAs were consistently consumed by the ants studied in this project and should be considered for pest ant control.

Harmful and Harmless Soil-Dwelling Fungi Indicate Microhabitat Suitability for Off-Host Ixodid Ticks.

Following blood meals or questing bouts, hard ticks (Ixodidae) must locate moist off-host microhabitats as refuge. Soil-dwelling fungi, including entomopathogenic Beauveria bassiana (Bb), thrive in moist microhabitats. Working with six species of ixodid ticks in olfactometer bioassays, we tested the hypothesis that ticks avoid Bb. Contrary to our prediction, nearly all ticks sought, rather than avoided, Bb-inoculated substrates. In further bioassays with female black-legged ticks, Ixodes scapularis, ticks oriented towards both harmful Bb and harmless soil-dwelling fungi, implying that fungi-regardless of their pathogenicity-signal habitat suitability to ticks. Only accessible Bb-inoculated substrate appealed to ticks, indicating that they sense Bb or its metabolites by contact chemoreception. Bb-inoculated substrate required ≥24 h of incubation before it appealed to ticks, suggesting that they respond to Bb metabolites rather than to Bb itself. Similarly, ticks responded to Bb-inoculated and incubated cellulose but not to sterile cellulose, indicating that Bb detection by ticks hinges on the Bb metabolism of cellulose. 2-Methylisoborneol-a common fungal metabolite with elevated presence in disturbed soils-strongly deterred ticks. Off-host ticks that avoid disturbed soil may lower their risk of physical injury. Synthetic 2-methylisoborneol could become a commercial tick repellent, provided its repellency extends to ticks in diverse taxa.

Blacklegged ticks, Ixodes scapularis, reduce predation risk by eavesdropping on communication signals of Formica oreas thatching ants.

Ticks spend most of their life inhabiting leaf litter and detritus where they are protected from sun but preyed upon by ants. Ants secrete chemical communication signals to coordinate group tasks such as nest defence. Ticks that avoid ant semiochemicals-as indicators of ant presence-would reduce predation risk by ants. We tested the hypotheses that: (i) chemical deposits from the thatching ant Formica oreas deter blacklegged ticks, Ixodes scapularis, (ii) deterrent semiochemicals originate from the ants' poison and/or Dufour's gland(s), and (iii) tick-deterrent semiochemicals serve as alarm-recruitment pheromone components in F. oreas. In two-choice olfactometer bioassays, filter paper soiled with ant chemical deposits significantly deterred female and male ticks. Poison and Dufour's gland extracts deterred ticks in combination but not alone. Gas chromatographic-mass spectrometric analyses of gland extracts revealed formic acid as the major constituent in the poison gland and eight hydrocarbons as constituents in the Dufour's gland. Synthetic formic acid and hydrocarbons deterred ticks only when combined. F. oreas workers sprayed both formic acid and hydrocarbons when distressed. A synthetic blend of these compounds elicited alarm-recruitment responses by F. oreas in behavioural bioassays. All results combined indicate that ticks eavesdrop on the ants' communication system.

Spectral sensitivity of click beetles (Coleoptera: Elateridae) and their responses to light stimuli in laboratory and field experiments.

With increasingly fewer insecticides registered to control the larvae of pest click beetles (Coleoptera: Elateridae), integrative beetle management, including pheromone- and light-based trapping of adult beetles, must be explored as an alternative strategy. Here, we analyzed the spectral sensitivity and color preference of 9 elaterids across 6 genera in electrophysiological recordings and in behavioral bioassays. In electroretinogram recordings (ERGs), dark-adapted beetles were exposed to narrow wavebands of light in 10-nm increments from 330 to 650 nm. All beetles proved most sensitive to green (515-538 nm) and ultraviolet (UV) light (~360 nm). In 4-choice bioassay arenas with 3 light emitting diodes (LEDs; green [525 nm], blue [470 nm], red [655 nm]) and a dark control as test stimuli, beetles discriminated between test stimuli, being preferentially attracted to green and blue LEDs. In field experiments, Vernon pitfall traps fitted with a green, blue or white LED captured significantly more male and female Agriotes lineatus and A. obscurus than dark control traps. When traps were baited with green or blue LEDs at light intensities that differed by 10-fold, the traps baited with higher light intensity lures captured numerically more beetles but trap catch data in accordance with light intensity did not differ statistically. Light-based trapping may be a viable tool for monitoring elaterid species known not to have pheromones.

Non-targeted metabolomics aids in sex pheromone identification: a proof-of-concept study with the triangulate cobweb spider, Steatoda triangulosa.

Targeted metabolomics has been widely used in pheromone research but may miss pheromone components in study organisms that produce pheromones in trace amount and/or lack bio-detectors (e.g., antennae) to readily locate them in complex samples. Here, we used non-targeted metabolomics-together with high-performance liquid chromatography-mass spectrometry (HPLC-MS), gas chromatography-MS, and behavioral bioassays-to unravel the sex pheromone of the triangulate cobweb spider, Steatoda triangulosa. A ternary blend of three contact pheromone components [N-4-methylvaleroyl-O-isobutyroyl-L-serine (5), N-3-methylbutyryl-O-isobutyroyl-L-serine (11), and N-3-methylbutyryl-O-butyroyl-L-serine (12)] elicited courtship by S. triangulosa males as effectively as female web extract. Hydrolysis of 5, 11 and 12 at the ester bond gave rise to two mate-attractant pheromone components [butyric acid (7) and isobutyric acid (8)] which attracted S. triangulosa males as effectively as female webs. Pheromone components 11 and 12 are reported in spiders for the first time, and were discovered only through the use of non-targeted metabolomics and GC-MS. All compounds resemble pheromone components previously identified in widow spiders. Our study provides impetus to apply non-targeted metabolomics for pheromone research in a wide range of animal taxa.

Widow spiders alter web architecture and attractiveness in response to same-sex competition for prey and mates, and predation risk.

Female-female competition in animals has rarely been studied. Responses of females that compete context-dependently for mates and prey, and seek safety from predators, are ideally studied with web-building spiders. Cobwebs possess unique sections for prey capture and safety, which can be quantified. We worked with Steaoda grossa females because their pheromone is known, and adjustments in response to mate competition could be measured. Females exposed to synthetic sex pheromone adjusted their webs, indicating a perception of intra-sexual competition via their sex pheromone. When females sequentially built their webs in settings of low and high intra-sexual competition, they adjusted their webs to increase prey capture and lower predation risk. In settings with strong mate competition, females deposited more contact pheromone components on their webs and accelerated their breakdown to mate-attractant pheromone components, essentially increasing their webs' attractiveness. We show that females respond to sexual, social and natural selection pressures originating from intra-sexual competition.

Synergistic attraction of Western black-legged ticks, Ixodes pacificus, to CO2 and odorant emissions from deer-associated microbes.

Foraging ticks reportedly exploit diverse cues to locate their hosts. Here, we tested the hypothesis that host-seeking Western black-legged ticks, Ixodes pacificus, and black-legged ticks, I. scapularis, respond to microbes dwelling in sebaceous gland secretions of white-tailed deer, Odocoileus virginianus, the ticks' preferred host. Using sterile wet cotton swabs, microbes were collected from the pelage of a sedated deer near forehead, preorbital, tarsal, metatarsal and interdigital glands. Swabs were plated on agar, and isolated microbes were identified by 16S rRNA amplicon sequencing. Of 31 microbial isolates tested in still-air olfactometers, 10 microbes induced positive arrestment responses by ticks, whereas 10 others were deterrent. Of the 10 microbes prompting arrestment by ticks, four microbes-including Bacillus aryabhattai (isolates A4)-also attracted ticks in moving-air Y-tube olfactometers. All four of these microbes emitted carbon dioxide and ammonia as well as volatile blends with overlapping blend constituents. The headspace volatile extract (HVE) of B. aryabhattai (HVE-A4) synergistically enhanced the attraction of I. pacificus to CO2. A synthetic blend of HVE-A4 headspace volatiles in combination with CO2 synergistically attracted more ticks than CO2 alone. Future research should aim to develop a least complex host volatile blend that is attractive to diverse tick taxa.

Stable flies sense and behaviorally respond to the polarization of light.

Insects use their polarization-sensitive photoreceptors in a variety of ecological contexts including host-foraging. Here, we investigated the effect of polarized light on host foraging by the blood-feeding stable fly, Stomoxys calcitrans, a pest of livestock. Electroretinogram recordings with chromatic adaptation demonstrated that the spectral sensitivity of stable flies resembles that of other calyptrate flies. Histological studies of the flies' compound eye revealed differences in microvillar arrangement of ommatidial types, assumed to be pale and yellow, with the yellow R7 and pale R8 photoreceptors having the greatest polarization sensitivity. In behavioural experiments, stable flies preferred to alight on horizontally polarized stimuli with a high degree of linear polarization. This preferential response disappeared when either ultraviolet (UV) or human-visible wavelengths were omitted from light stimuli. Removing specific wavelength bands further revealed that the combination of UV (330-400 nm) and blue (400-525 nm) wavelength bands was sufficient to enable polarized light discrimination by flies. These findings enhance our understanding of polarization vision and foraging behavior among hematophagous insects and should inform future trap designs.

Characterization of New Defensin Antimicrobial Peptides and Their Expression in Bed Bugs in Response to Bacterial Ingestion and Injection.

Common bed bugs, Cimex lectularius, can carry, but do not transmit, pathogens to the vertebrate hosts on which they feed. Some components of the innate immune system of bed bugs, such as antimicrobial peptides (AMPs), eliminate the pathogens. Here, we determined the molecular characteristics, structural properties, and phylogenetic relatedness of two new defensins (CL-defensin1 (XP_024085718.1), CL-defensin2 (XP_014240919.1)), and two new defensin isoforms (CL-defensin3a (XP_014240918.1), CL-defensin3b (XP_024083729.1)). The complete amino acid sequences of CL-defensin1, CL-defensin2, CL-defensin3a, and CL-defensin3b are strongly conserved, with only minor differences in their signal and pro-peptide regions. We used a combination of comparative transcriptomics and real-time quantitative PCR to evaluate the expression of these defensins in the midguts and the rest of the body of insects that had been injected with bacteria or had ingested blood containing the Gram-positive (Gr+) bacterium Bacillus subtilis and the Gram-negative (Gr-) bacterium Escherichia coli. We demonstrate, for the first time, sex-specific and immunization mode-specific upregulation of bed bug defensins in response to injection or ingestion of Gr+ or Gr- bacteria. Understanding the components, such as these defensins, of the bed bugs' innate immune systems in response to pathogens may help unravel why bed bugs do not transmit pathogens to vertebrates.

Origin, structure and functional transition of sex pheromone components in a false widow spider.

Female web-building spiders disseminate pheromone from their webs that attracts mate-seeking males and deposit contact pheromone on their webs that induces courtship by males upon arrival. The source of contact and mate attractant pheromone components, and the potential ability of females to adjust their web's attractiveness, have remained elusive. Here, we report three new contact pheromone components produced by female false black widow spiders, Steatoda grossa: N-4-methylvaleroyl-O-butyroyl-L-serine, N-4-methylvaleroyl-O-isobutyroyl-L-serine and N-4-methylvaleroyl-O-hexanoyl-L-serine. The compounds originate from the posterior aggregate silk gland, induce courtship by males, and web pH-dependently hydrolyse at the carboxylic-ester bond, giving rise to three corresponding carboxylic acids that attract males. A carboxyl ester hydrolase (CEH) is present on webs and likely mediates the functional transition of contact sex pheromone components to the carboxylic acid mate attractant pheromone components. As CEH activity is pH-dependent, and female spiders can manipulate their silk's pH, they might also actively adjust their webs' attractiveness.

Seasonal and Diel Communication Periods of Sympatric Pest Limonius Click Beetle Species (Coleoptera: Elateridae) in Western Canada.

In western North America, sympatric Limonius click beetle species produce limoniic acid [(E)-4-ethyloct-4-enoic acid] as a sex pheromone component (L. canus (LeConte), L. californicus (Mannerheim)) or respond to it as a sex attractant (L. infuscatus (Motschulsky)). We tested the hypothesis that these three congeners maintain species-specificity of sexual communication through nonoverlapping seasonal occurrence and/or contrasting diel periodicity of sexual communication. Using capture times of beetles in pheromone-baited traps as a proxy for sexual communication periods, our data show that L. canus and L. californicus have seasonally distinct communication periods. Most L. canus males (>90%) were captured in April and most L. californicus males (>95%) were captured in May/June/July. As almost exclusively L. infuscatus males were captured in two separate 24-hr trapping studies, with data recordings every hour, it remains inconclusive whether the three Limonius congeners communicate at different times of the day. Males of L. infuscatus responded to pheromone lures only during daytime hours and during the warmest period each day. Captures of L. infuscatus overlapping with those of L. canus in April and those of L. californicus in May/June imply the presence of reproductive isolating mechanisms other than seasonal separation of sexual communication periods.

Identification of the Major Sex Pheromone Component of the Click Beetle Agriotes ferrugineipennis.

Synthetic sex pheromone lures are useful tools to monitor and control populations of adult click beetles (Coleoptera: Elateridae). However, sex pheromones for Agriotes click beetle species native to North America have yet to be identified. Here we report the identification and field testing of the sex pheromone of Agriotes ferrugineipennis. Headspace volatiles from female beetles were collected on Porapak Q, and aliquots of Porapak extract were analyzed by gas chromatographic-electroantennographic detection (GC-EAD) and GC-mass spectrometry. 7-Methyloctyl 7-methyloctanoate (7Me7Me) emitted by females was more abundant and elicited much stronger responses from male antennae than the aldehydes octanal and nonanal and the ketone 6,10,14-trimethyl-2-pentadecanone. In a field experiment, captures of A. ferrugineipennis males in traps baited with candidate pheromone components exceeded those of unbaited control traps, on average by nearly 1,200 times. Neither the ketone nor the aldehydes as lure constituents appeared to alter captures of males in 7Me7Me-baited traps. We conclude that 7Me7Me is the major, and possibly the only, sex attractant pheromone component of female A. ferrugineipennis.

Identification of the Trail Pheromone of the Pavement Ant Tetramorium immigrans (Hymenoptera: Formicidae).

Four species of Tetramorium pavement ants are known to guide foraging activities of nestmates via trail pheromones secreted from the poison gland of worker ants, but the trail pheromone of T. immigrans is unknown. Our objectives were to (1) determine whether poison gland extract of T. immigrans workers induces trail-following behavior of nestmates, (2) identify the trail pheromone, and (3) test whether synthetic trail pheromone induces trail-following behavior of workers. In laboratory no-choice bioassays, ants followed poison-gland-extract trails farther than they followed whole-body-extract trails or solvent-control trails. Gas chromatographic-electroantennographic detection (GC-EAD) analyses of poison gland extract revealed a single candidate pheromone component (CPC) that elicited responses from worker ant antennae. The CPC mass spectrum indicated, and an authentic standard confirmed, that the CPC was methyl 2-methoxy-6-methylbenzoate (MMMB). In further laboratory no-choice bioassays, ants followed poison-gland-extract trails (tested at 1 ant equivalent) and synthetic MMMB trails (tested at 0.35 ant equivalents) equally far, indicating that MMMB is the single-component trail pheromone of T. immigrans. Moreover, in laboratory two-choice bioassays, ants followed MMMB trails ~ 21-times farther than solvent-control trails. In field settings, when T. immigrans colonies were offered a choice between two paper strips treated with a synthetic MMMB trail or a solvent-control trail, each leading to an apple bait, the MMMB trails efficiently recruited nestmates to baits.

Time- and tissue-specific antimicrobial activity of the common bed bug in response to blood feeding and immune activation by bacterial injection.

Unlike almost all hematophagous insects, common bed bugs, Cimex lectularius, are not known to transmit pathogens to humans. To help unravel the reasons for their lack of vector competence, we studied the time- and tissue-dependent expression of innate immune factors after blood feeding or immune activation through the intrathoracic injection of bacteria. We used minimum inhibitory concentration (MIC1) bioassays and the Kirby-Bauer protocol to evaluate antimicrobial peptide (AMP2) activity in tissue extracts from the midguts or 'rest of body' (RoB3) tissues (containing hemolymph and fat body AMPs) against Gram-positive and Gram-negative bacteria. We compared AMP activity between blood-fed female bed bugs and yellow fever mosquitoes, Aedes aegypti and determined how female and male bed bugs respond to immune challenges, and how long AMP gene expression remains elevated in bed bugs following a blood meal. Blood meal-induced AMP activity is 4-fold stronger in female bed bugs than in female mosquitoes. Male bed bugs have elevated AMP activity within 8 h of a blood meal or an intrathoracic injection with bacteria, with the strongest activity expressed in RoB tissue 24 h after the immune challenge. Female bed bugs have a stronger immune response than males within 24 h of a blood meal. The effects of blood meal-induced elevated AMP activity lasts longer against the Gram-positive bacterium, Bacillus subtilis, than against the Gram-negative bacterium Escherichia coli. Unravelling the specific immune pathways that are activated in the bed bugs' immune responses and identifying the bed bug-unique AMPs might help determine why these insects are not vectors of human parasites.

All sugars ain't sweet: selection of particular mono-, di- and trisaccharides by western carpenter ants and European fire ants.

Ants select sustained carbohydrate resources, such as aphid honeydew, based on many factors including sugar type, volume and concentration. We tested the hypotheses (H1-H3) that western carpenter ants, Camponotus modoc, seek honeydew excretions from Cinara splendens aphids based solely on the presence of sugar constituents (H1), prefer sugar solutions containing aphid-specific sugars (H2) and preferentially seek sugar solutions with higher sugar content (H3). We further tested the hypothesis (H4) that workers of both Ca. modoc and European fire ants, Myrmica rubra, selectively consume particular mono-, di- and trisaccharides. In choice bioassays with entire ant colonies, sugar constituents in honeydew (but not aphid-specific sugar) as well as sugar concentration affected foraging decisions by Ca. modoc. Both Ca. modoc and M. rubra foragers preferred fructose to other monosaccharides (xylose, glucose) and sucrose to other disaccharides (maltose, melibiose, trehalose). Conversely, when offered a choice between the aphid-specific trisaccharides raffinose and melezitose, Ca. modoc and M. rubra favoured raffinose and melezitose, respectively. Testing the favourite mono-, di- and trisaccharide head-to-head, both ant species favoured sucrose. While both sugar type and sugar concentration are the ultimate cause for consumption by foraging ants, strong recruitment of nest-mates to superior sources is probably the major proximate cause.