Differential development and survival of Blattella asahinai and Blattella germanica (Blattodea: Ectobiidae) at six constant temperatures.

The Asian cockroach, Blattella asahinai Mizukubo, is a peridomestic nuisance pest in the southeastern United States. Blattella asahinai is the closest relative to Blattella germanica (L.), the German cockroach, one of the most prolific and widespread domestic pests. Because these two species live in different habitats, they are expected to have differential development patterns reflecting environmental adaptations. Development of B. asahinai and B. germanica cockroach nymphs were observed at six constant temperatures ranging from 10 to 35 °C. At 10 °C and 15 °C, all nymphs died in the first instar, but B. germanica nymphs survived longer (10 °C: 12.9 d; 15 °C: 42.9 d) than B. asahinai nymphs (10 °C 8.2 d; 15 °C 18.4 d) at both temperatures. At 20 °C, 25 °C, and 30 °C, B. asahinai consistently had more instars and longer stadia than B. germanica. At 35 °C, only B. germanica was able to complete nymphal development; cannibalism among B. asahinai nymphs during molting was often observed at this temperature. The results for B. asahinai corroborated previously estimated growth patterns. The lower nymphal development threshold was 14.1 °C for B. germanica and 13.7 °C for B. asahinai. Comparing the development of B. germanica directly with its closest relative reveals specific physiological adaptations that B. germanica has developed for the indoor biome.

Potential of the oothecal parasitoid Aprostocetus hagenowii (Hymenoptera: Eulophidae) as a biological control agent for the Turkestan cockroach (Blattodea: Blattidae).

The Turkestan cockroach, Blatta lateralis (Walker), is a peridomestic pest of growing concern in the US Southwest. The parasitoid Aprostocetus hagenowii (Ratzburg) is used in IPM programs targeting other blattid cockroach species and may aid in B. lateralis suppression. Information about the ability of A. hagenowii to parasitize B. lateralis is lacking. A no-choice host-switching experiment was used to test A. hagenowii acceptance of B. lateralis oothecae, and a multigenerational no-choice experiment was used to determine the suitability of B. lateralis as a host for A. hagenowii over several months of rearing. Periplaneta americana (L.) (Blattodea: Blattidae), the preferred host of A. hagenowii, and Blatta orientalis L., a known host and relative of B. lateralis, were used for comparison. Development time was similar among hosts and generations (P > 0.05). Parasitism success and proportion of female progeny declined significantly with subsequent generations on both Blatta spp. (parasitism success: χ2 = 14.916; df = 2; P = 0.001; proportion female: H = 6.364; df = 2; P = 0.041). These results suggest that A. hagenowii may initially aid in suppression of B. lateralis, but an overall decline in fitness will require repeated releases or provisioning of P. americana oothecae. Development of a strain more suitable for B. lateralis control may be possible via selection from laboratory strains or through use of wild A. hagenowii from areas where B. lateralis is present.

The prevalence of Wolbachia in multiple cockroach species and its implication for urban insect management.

Cockroach management relies heavily on the use of conventional insecticides in urban settings, which no longer provide the anticipated level of control. Knowledge of cockroach endosymbionts, like Wolbachia, might provide novel avenues for control. Therefore, we screened 16 cockroach species belonging to 3 families (Ectobiidae, Blattidae, and Blaberidae) for the presence of Wolbachia. We mapped the evolution of Wolbachia-cockroach relationships based on maximum likelihood phylogeny and phylogenetic species clustering on a multi-loci sequence dataset (i.e., coxA, virD4, hcpA, and gatB) of Wolbachia genes. We confirmed the previous report of Wolbachia in 1 Ectobiid species; Supella longipalpa (Fab.), and detected the presence of Wolbachia in 2 Ectobiid species; Balta notulata (Stål) and Pseudomops septentrionalis Hebard, and 1 Blaberid species; Gromphadorhina portentosa (Schaum). All cockroach-associated Wolbachia herein detected were clustered with the ancestor of F clade Wolbachia of Cimex lectularius L. (bed bugs). Since Wolbachia provision C. lectularius with biotin vitamins that confer reproductive fitness, we screened the cockroach-associated Wolbachia for the presence of biotin genes. In toto, our results reveal 2 important findings: (i) Wolbachia is relatively uncommon among cockroach species infecting about 25% of species investigated, and (ii) cockroach-associated Wolbachia have biotin genes that likely provide nutritional benefits to their hosts. Thus, we discuss the potential of exploring Wolbachia as a tool for urban insect management.

Flying on empty: reduced mitochondrial function and flight capacity in food-deprived monarch butterflies.

Mitochondrial function is fundamental to organismal performance, health and fitness - especially during energetically challenging events, such as migration. With this investigation, we evaluated mitochondrial sensitivity to ecologically relevant stressors. We focused on an iconic migrant, the North American monarch butterfly (Danaus plexippus), and examined the effects of two stressors: 7 days of food deprivation and infection by the protozoan parasite Ophryocystis elektroscirrha (known to reduce survival and flight performance). We measured whole-animal resting metabolic rate (RMR) and peak flight metabolic rate, and mitochondrial respiration of isolated mitochondria from the flight muscles. Food deprivation reduced mass-independent RMR and peak flight metabolic rate, whereas infection did not. Fed monarchs used mainly lipids in flight (respiratory quotient 0.73), but the respiratory quotient dropped in food-deprived individuals, possibly indicating switching to alternative energy sources, such as ketone bodies. Food deprivation decreased mitochondrial maximum oxygen consumption but not basal respiration, resulting in lower respiratory control ratio (RCR). Furthermore, food deprivation decreased mitochondrial complex III activity, but increased complex IV activity. Infection did not result in any changes in these mitochondrial variables. Mitochondrial maximum respiration rate correlated positively with mass-independent RMR and flight metabolic rate, suggesting a link between mitochondria and whole-animal performance. In conclusion, low food availability negatively affects mitochondrial function and flight performance, with potential implications for migration, fitness and population dynamics. Although previous studies have reported poor flight performance in infected monarchs, we found no differences in physiological performance, suggesting that reduced flight capacity may be due to structural differences or low energy stores.

Profiles of telomeric repeats in Insecta reveal diverse forms of telomeric motifs in Hymenopterans.

Telomeres consist of highly conserved simple tandem telomeric repeat motif (TRM): (TTAGG)n in arthropods, (TTAGGG)n in vertebrates, and (TTTAGGG)n in most plants. TRM can be detected from chromosome-level assembly, which typically requires long-read sequencing data. To take advantage of short-read data, we developed an ultra-fast Telomeric Repeats Identification Pipeline and evaluated its performance on 91 species. With proven accuracy, we applied Telomeric Repeats Identification Pipeline in 129 insect species, using 7 Tbp of short-read sequences. We confirmed (TTAGG)n as the TRM in 19 orders, suggesting it is the ancestral form in insects. Systematic profiling in Hymenopterans revealed a diverse range of TRMs, including the canonical 5-bp TTAGG (bees, ants, and basal sawflies), three independent losses of tandem repeat form TRM (Ichneumonoids, hunting wasps, and gall-forming wasps), and most interestingly, a common 8-bp (TTATTGGG)n in Chalcid wasps with two 9-bp variants in the miniature wasp (TTACTTGGG) and fig wasps (TTATTGGGG). Our results identified extraordinary evolutionary fluidity of Hymenopteran TRMs, and rapid evolution of TRM and repeat abundance at all evolutionary scales, providing novel insights into telomere evolution.

Why Do Insects Close Their Spiracles? A Meta-Analytic Evaluation of the Adaptive Hypothesis of Discontinuous Gas Exchange in Insects.

The earliest description of the discontinuous gas exchange cycle (DGC) in lepidopterous insects supported the hypothesis that the DGC serves to reduce water loss (hygric hypothesis) and facilitate gaseous exchange in hyperoxia/hypoxia (chthonic hypothesis). With technological advances, other insect orders were investigated, and both hypotheses were questioned. Thus, we conducted a meta-analysis to evaluate the merit of both hypotheses. This included 46 insect species in 24 families across nine orders. We also quantified the percent change in metabolic rates per °C change of temperature during the DGC. The DGC reduced water loss (-3.27 ± 0.88; estimate ± 95% confidence limits [95% CI]; p < 0.0001) in insects. However, the DGC does not favor gaseous exchange in hyperoxia (0.21 ± 0.25 [estimate ± 95% CI]; p = 0.12) nor hypoxia, but did favor gaseous exchange in normoxia (0.27 ± 0.26 [estimate ± 95% CI]; p = 0.04). After accounting for variation associated with order, family, and species, a phylogenetic model reflected that metabolic rate exhibited a significant, non-zero increase of 8.13% (± 3.48 95% CI; p < 0.0001) per °C increase in temperature. These data represent the first meta-analytic attempt to resolve the controversies surrounding the merit of adaptive hypotheses in insects.

A Review of Alternative Management Tactics Employed for the Control of Various Cockroach Species (Order: Blattodea) in the USA.

Effective control of domestic and peridomestic cockroaches requires integrated approaches that emphasize concurrent use of chemicals with alternative control tactics. An integrated pest management (IPM) approach is particularly justified in environments where satisfactory cockroach control cannot be achieved due to multiple factors including development of insecticide aversion and resistance in some cockroach species, and poor sanitation or structural issues that foster infestations. While a flurry of research effort has been devoted to study alternative tactics for cockroach control, only a few of them have been evaluated in the context of IPM programs. This review focuses on examining studies on alternative tactics that are proven efficacious, economical, and logistically feasible for their inclusion in IPM programs for important domestic and peridomestic cockroaches in the USA. Management programs that educate the public on cockroach biology, behavior, and the importance of sanitation; use of traps to monitor infestation levels; apply targeted low impact insecticides such as baits, have demonstrated a greater success for effective and sustainable control of cockroaches when compared to an insecticide-only approach. Incorporation of other alternative control methods to IPM programs will require more applied research that validates their use in real-world scenarios and demonstrates their cost-effectiveness.

Comparative Cutaneous Water Loss and Desiccation Tolerance of Four Solenopsis spp. (Hymenoptera: Formicidae) in the Southeastern United States.

The high surface area to volume ratio of terrestrial insects makes them highly susceptible to desiccation mainly through the cuticle. Cuticular permeability (CP) is usually the most important factor limiting water loss in terrestrial insects. Water loss rate, percentage of total body water (%TBW) content, CP, and desiccation tolerance were investigated in workers of four Solenopsis species in the southeastern USA. We hypothesized that tropical/subtropical ants (S. invicta and S. geminata) will have lower CP values and tolerate higher levels of desiccation than temperate ants (S. richteri and S. invicta × S. richteri). The %TBW content was similar among species. Solenopsis invicta had a 1.3-fold and 1.1-fold lower CP value than S. invicta × S. richteri and S. richteri, respectively. Solenopsis geminata had a 1.3-fold lower CP value than S. invicta × S. richteri, and a 1.2-fold lower CP value than S. richteri. The LT50 values (lethal time to kill 50% of the population) ranged from 1.5 h (small S. geminata) to 8.5 h (large S. invicta). Desiccation tolerance ranged between 36 and 50 %TBW lost at death and was not related to a species' location of origin. This study is the first report of water relations of S. invicta × S. richteri. It demonstrates that desiccation stress differentially can affect the survival of different Solenopsis species and implies that environmental stress can affect the distribution of these species in the southeastern USA.

Water Loss and Desiccation Tolerance of the Two Yearly Generations of Adult and Nymphal Kudzu Bugs, Megacopta cribraria (Hemiptera: Plataspidae).

Water loss rate, percentage total body water content (%TBW), cuticular permeability (CP), and desiccation tolerance were investigated in adult and immature stages of the invasive kudzu bug, Megacopta cribraria (Fab.) (Hemiptera: Plataspidae), a serious soybean pest and an urban nuisance. Adults and all five nymphal instars were weighed prior to and 2, 4, 6, 8, 10, and 24 h after desiccated at 30 ± 1°C and 0-2% RH. Both % initial mass and %TBW loss increased linearly with time of desiccation. Rates of loss ranged from approximately 1-7%/h. Mortality occurred at 10 h after desiccation. Desiccation tolerance (%TBW lost at death) ranged between 25.6% for first-generation adult females and 75% for first-generation fifth-instar nymphs. First-generation first-instar nymphs had significantly greater %TBW (88.9%) than the other generations and instars, whereas second-generation fifth instars had the lowest %TBW (62.4%). The CP value of first-generation adult females (12.3 ± 1.6 µg cm-1 h-1 mmHg-1) was the greatest across generations. First-generation first instars had the greatest mass loss (111.11 mg/g) among all instars and generations, whereas overwintered second-generation adult females had the lowest mass loss (18.39) across generations. This study demonstrated that desiccation stress differentially affected the survival of adult and nymphal kudzu bugs and may imply that environmental stress can affect the relative abundance of this species in the fields and around homes.

Innate immunity of Florida cane toads: how dispersal has affected physiological responses to LPS.

Physiological tradeoffs occur in organisms coping with their environments, which are likely to increase as populations reach peripheries of established ranges. Invasive species offer opportunities to study tradeoffs that occur, with many hypotheses focusing on how immune responses vary during dispersal. The cane toad (Rhinella marina) is a well-known invasive species. Populations near the expanding edge of the Australian invasion have altered immune responses compared to toads from longer-established core populations, although this has not been well-documented for Florida populations. In this study, cane toads from a northern edge [New Port Richey (NPR)] and southern core (Miami) population in Florida were collected and injected with lipopolysaccharide (LPS) to compare immune responses. Core population individuals injected with LPS showed greater metabolic increases compared to their baseline rates that were higher compared to those from the edge population. In addition, LPS-injected core individuals had different circulating leukocyte profiles compared to saline-injected cane toads while edge individuals did not. There was a significant interaction between plasma bacteria-killing capability (BKA) and treatment, such that BKA decreased with time in saline compared to LPS-injected individuals, and saline-injected toads from the edge population had lower BKA compared to LPS-injected edge toads at 20 h post-injection. There was also a significant interaction between location and time on circulating corticosterone (CORT) levels following injections with saline or LPS, with CORT decreasing more with time in core population toads. The differential CORT response indicates that differential stress responses contribute to the tradeoffs observed with immunity and dispersal.

Instar Determination of Blattella asahinai (Blattodea: Ectobiidae) From Digital Measurements of the Pronotum Using Gaussian Mixture Modeling and the Number of Cercal Annuli.

The Asian cockroach, Blattella asahinai Mizukubo, has expanded its range throughout the southeastern United States since its introduction into Florida. Unlike its closest relative, the German cockroach, Blattella germanica (L.), B. asahinai lives outdoors and can fly. There is little information on the biology and development of B. asahinai, including the number of instars during nymphal development. To estimate the number of instars of B. asahinai, nymphs were photographed, sexed, and the lengths and widths of their pronota were measured digitally. The number of instars of B. asahinai was estimated using Gaussian mixture models with the pronotal data. The most probable model and its clusters were selected to assign individuals to an instar. Instars were also determined by counting the number of cercal annuli of nymphs. Both clustering and cercal annuli indicated that B. asahinai most frequently had six instars when reared at 30°C. Growth did not strictly follow the Brooks-Dyar Rule, because nymphs had different numbers of instars and different growth patterns. Although Gaussian mixture models are not efficient for field sampling experiments, digital measurements may provide a way to estimate instars with live specimens in development studies without handling the animals in a way that may alter growth.

Temperature-Mediated Variations in Behavior and Mortality Caused by Non-Repellent Insecticides in Subterranean Termites (Blattodea: Rhinotermitidae).

Behavioral symptoms and mortality associated with intoxication with insecticides fipronil and indoxacarb were determined in field-collected eastern subterranean termites, Reticulitermes flavipes (Kollar), and Formosan subterranean termites, Coptotermes formosanus Shiraki. Behaviors and mortality were evaluated at three temperatures (16, 22, and 28 °C) and three concentrations of fipronil (0.5, 1, and 5 ppm) and indoxacarb (50, 75, and 100 ppm). LT50 (median lethal time to kill 50% of the termites) values declined with increasing concentrations and temperatures for both fipronil-exposed eastern and Formosan subterranean termites, whereas these values were not always the highest at 16 °C for indoxacarb-treated termites. The greatest change (reduction) in LT50 values occurred for both species between 16 and 22 °C at the lowest concentration of each insecticide. Intoxication and moribundity were the most frequently observed behaviors for fipronil-exposed termites, whereas intoxication, ataxia, and moribundity were observed for most concentration and temperature combinations for indoxacarb-exposed termites. The inherent toxicity of fipronil was higher than that of indoxacarb. The higher presence and duration of intoxication behaviors may positively affect the performance of indoxacarb against subterranean termite colonies.

Repellency and Laboratory Performance of Selected Insecticides to Field-Collected Insecticide Resistant German Cockroaches (Blattodea: Ectobiidae).

German cockroaches, Blattella germanica (L.) (Blattodea: Ectobiidae), are important indoor insect pests and remain difficult to control because of their ability to develop resistance to insecticides. The toxicity, resistance levels, repellency, and performance index (PI) value of five formulated insecticides (permethrin, chlorpyrifos, propoxur, imidacloprid, and fipronil) were determined for adult males of seven strains of the German cockroach: a laboratory-reared susceptible strain (S) and six field-collected strains (B, D, E, G, H, and I). Propoxur was generally the most toxic insecticide to all strains using continuous exposure methods; however, using Ebeling choice box methods, chlorpyrifos, and fipronil were most toxic. In both continuous exposure and Ebeling choice box tests, the field-collected strains were generally most resistant to permethrin among the five insecticides. The greatest increase in resistance ratios between the two exposure methods was for permethrin and propoxur. Permethrin was the most repellent insecticide against all but two strains (D and H). PI values reached 100 (no repellency and complete mortality) for the susceptible strain against only chlorpyrifos and fipronil. Only fipronil resulted in a PI value of 100 for any of the field-collected strains. Continuous exposure and Ebeling choice box resistance ratios were correlated, and both were correlated with previously reported resistance ratios (based on LD50 values). Choice box repellency, however, was only correlated with LD50 resistance ratios. PIMax was negatively correlated with all measures of resistance ratios.

Cuticular hydrocarbon chemistry, an important factor shaping the current distribution pattern of the imported fire ants in the USA.

Two sibling species, Solenopsis richteri and S. invicta, were both introduced into the southern USA from South America in the early 20th century. Today, S. richteri occupies higher latitudes and colder areas, while S. invicta occupies lower latitudes. Between the distributions of the two species, there is a large area of viable hybrid (S. richteri × S. invicta) populations. This study aimed to characterize the forces driving this distribution pattern and the underlying mechanisms. Cuticular hydrocarbons (CHCs) of freshly killed workers of S. invicta, hybrids, and S. richteri were removed using hexane. Both intact and CHCs-extracted workers were subjected to a constant rate of increasing temperature from 10 to 60 °C to obtain relative water loss and the water loss transition temperature (Tc-ant). Mass loss and Tc-ant were both significantly increased with CHCs removal. We then examined the CHC composition of three species. CHC profiles of S. richteri are characterized by significant amounts of short-chain (C23-C27) saturated and unsaturated hydrocarbons. In contrast, profiles of S. invicta consist primarily of long-chain (C27-C29) saturated hydrocarbons; unsaturated alkenes are completely lacking. Hybrid fire ants show intermediate profiles of the two parent species. We measured the melting point (Tm) and water-loss transition temperature of CHC blends (Tc-CHC) of different ant species colonies using differential scanning calorimetry (DSC) and an artificial membrane system, respectively. There were 3-5 Tms of each CHCs sample of different ant colonies due to their complex chemistry. The highest Tms (Tm-maxs) of CHCs samples from S. invicta and the hybrid were significantly higher than that from S. richteri. The correlation between Tc-CHC and Tm-max obtained from the same CHCs sample was highly significant. These results reveal that species having higher Tc and Tm-max retain more water under relatively higher temperature, and consequently are able to occupy warmer environments. We conclude that CHC chemistry plays a role in shaping current distribution patterns of S. richteri, S. invicta and their hybrid in the United States.

Insecticide Resistance of Several Field-Collected German Cockroach (Dictyoptera: Blattellidae) Strains.

German cockroaches, Blattella germanica (L.), remain one of the most difficult indoor insect species to control because of its ability to develop resistance to insecticides. The toxicity and resistance levels of five technical-grade insecticides (permethrin, chlorpyrifos, propoxur, imidacloprid, and fipronil) were determined for adult males of seven strains of the German cockroach, a laboratory-reared susceptible strain (S) and six field-collected strains (B, D, E, G, H, and I). Using topical application methods, fipronil was the most toxic insecticide to all seven strains. The LD50 values of fipronil in the susceptible strain (S) and the field-collected strains B, D, E, G, H, and I were 1.33, 2.62, 11.53, 5.07, 7.66, 5.15, and 10.15 ng/insect, respectively. The field-collected strains were most resistant to permethrin among the five insecticides, except for strain H. The resistance ratios of strains B, D, E, G, and I to permethrin were 31.8, 37.3, 51.9, 34.9, and 37.5, respectively. With a resistance ratio of 6.4, the field-collected strain H was most resistant to chlorpyrifos. The field-collected strains were not significantly resistant to propoxur. Strains B, H, and I were not significantly resistant to imidacloprid when compared with the susceptible strain. Based on the different resistance ratios for each insecticide, we conclude that there are high rates of insecticide resistance in German cockroaches from Franklin County, NC, and that the field-collected strains most likely had different treatment histories.

Temperature-Dependent Development and Thermal Sensitivity of Blaptica dubia (Blattodea: Blaberidae).

Temperature-dependent development of nymphs of the Dubia cockroach, Blaptica dubia Serville, was described using constant-temperature data collected from laboratory experiments at 15, 20, 25, 30, 35, and 40 °C. Simple linear regression models were developed based on the data from each instar. Degree-days required to complete a particular life stage were estimated as 457.5, 668, 1,031, 1,317, 1,515, 1,429, and 2,071 for instars 1-7, respectively. Nymphs developed to adulthood at 20, 25, and 30 °C, but all died before developing into fifth instar at 35 °C and into second instar at 40 °C. Critical thermal maximum (CTMax) and critical thermal minimum (CTMin) of B. dubia were also measured for each instar. CTMax ranged from 44.8 to 49.9 °C for fourth and second instars, respectively. CTMin ranged from approximately -2 °C for seventh instar to - 3.1 °C for second instar. There was no relationship between body mass (instar) and CTMax; however, CTMin was positively correlated with body mass. These results could be used to control the development rate of B. dubia and adjust the optimal rearing temperature for B. dubia in a given situation.

Reduced innate immunity of Cuban Treefrogs at leading edge of range expansion.

During geographic range expansion, populations of non-indigenous species at the invasion front may benefit from directing resources away from immune defense. To test this hypothesis, we investigated the strength of two innate immune components in populations of invasive Cuban Treefrogs (Osteopilus septentrionalis) in a long-colonized area (core region) and at the invasion front (leading-edge region). First, we compared the region-specific metabolic response of frogs injected with an endotoxin that induces systemic inflammation (lipopolysaccharide, LPS) to sham-injected control frogs pooled from both regions. Males and females were analyzed independently because we detected a sex-related difference in mass-independent metabolism of control frogs, with males exhibiting a significantly higher metabolic rate (F1, 21  = 29.02, P < 0.001) than females. We observed a significantly higher metabolic rate in LPS-injected core frogs compared with control frogs for both males (P = 0.041) and females (P = 0.007). Conversely, in leading-edge populations, there was no significant difference in the metabolic rate of LPS-injected and control frogs (males, P  = 0.195; females, P  = 0.132). Second, we directly compared bacterial killing ability of frog blood plasma between regions. Bactericidal ability of plasma was significantly greater in frogs from the core region in comparison with those at the leading edge (F1, 26   = 28.67, P < 0.001). For both immune components that we examined, populations from the core exhibited stronger immune responses. Our findings support hypotheses predicting an inverse relationship between immunity and range expansion.

Estimating the critical thermal maximum (CTmax) of bed bugs, Cimex lectularius: Comparing thermolimit respirometry with traditional visual methods.

Evaluating the critical thermal maximum (CTmax) in insects has provided a number of challenges. Visual observations of endpoints (onset of spasms, loss of righting response, etc.) can be difficult to measure consistently, especially with smaller insects. To resolve this problem, Lighton and Turner (2004) developed a new technique: thermolimit respirometry (TLR). TLR combines real time measurements of both metabolism (V·CO2) and activity to provide two independent, objective measures of CTmax. However, several questions still remain regarding the precision of TLR and how accurate it is in relation to traditional methods. Therefore, we evaluated CTmax of bed bugs using both traditional (visual) methods and TLR at three important metabolic periods following feeding (1d, 9d, and 21d). Both methods provided similar estimates of CTmax, although traditional methods produced consistently lower values (0.7-1°C lower than TLR). Despite similar levels of precision, TLR provided a more complete profile of thermal tolerance, describing changes in metabolism and activity leading up to the CTmax, not available through traditional methods. In addition, feeding status had a significant effect on bed bug CTmax, with bed bugs starved 9d (45.19[±0.20]°C) having the greatest thermal tolerance, followed by bed bugs starved 1d (44.64[±0.28]°C), and finally bed bugs starved 21d (44.12[±0.28]°C). Accuracy of traditional visual methods in relation to TLR is highly dependent on the selected endpoint; however, when performed correctly, both methods provide precise, accurate, and reliable estimations of CTmax.

Effects of Starvation on Deltamethrin Tolerance in Bed Bugs, Cimex lectularius L. (Hemiptera: Cimicidae).

Bed bugs, Cimex lectularius L., are a major pest in the urban environment. Their presence often results in physical, psychological, and financial distress of homeowners and apartment dwellers. Although many insecticide bioassays have been performed on this pest, little attention has been paid to bed bug feeding status, which is closely linked to metabolism, molting, and mass. Therefore, we evaluated the toxicity of topically applied deltamethrin on insecticide susceptible adult male bed bugs fed 2 d, 9 d, and 21 d prior to testing. When toxicity was evaluated on a "per-bug" basis, there was no difference between 2 d [LD50 = 0.498 (0.316 - 0.692) ng·bug(-1)] and 9 d [LD50 = 0.572 (0.436 - 0.724) ng·bug(-1)] starved bugs, while 21 d starved bugs had a significantly lower LD50 [0.221 (0.075 - 0.386) ng·bug(-1)]. When toxicity was evaluated in terms of body mass, 9 d starved bugs had the highest LD50 values [0.138 (0.102 - 0.176) ng·mg(-1)], followed by 2 d starved bugs [0.095 (0.060 - 0.134) ng·mg(-1)], and then 21 d starved bugs [0.058 (0.019-0.102) ng·mg(-)¹]; the LD50 values of 2 d and 9 d starved bugs were significantly different from 21 d starved bugs. These results indicate that feeding status plays an important role in the toxicity of deltamethrin. In addition, the lack of differences between 2 d and 9 d starved bugs indicate that the blood meal itself has little impact on tolerance, but rather it is some physiological change following feeding that confers increased tolerance to bed bugs.

Effects of starvation and molting on the metabolic rate of the bed bug (Cimex lectularius L.).

The bed bug (Cimex lectularius L.) is a common hematophagous pest in the urban environment and is capable of surviving extended periods of starvation. However, the relationship between starvation and metabolism in bed bugs is not well understood. To better understand this relationship, we measured the metabolism of all life stages for >900 h after feeding (starvation) using closed-system respirometry. Measurements were made around molting for the immature life stages, which occurs only after a blood meal. In addition, both mated and unmated adults were measured. Starvation and molting had significant effects on the metabolism of the bed bug. Mass-specific metabolic rate (V(O2); mL g(-1) h(-1)) declined in a curvilinear fashion with the period of starvation for adults and with the postmolting period for immature bed bugs (used to standardize all immature life stages). A standard curve was developed to depict the generalized pattern of metabolic decline observed in all life stages that molted. Individual metabolic comparisons among life stages that molted revealed some differences in metabolic rate between unmated males and females. In addition, the mass scaling coefficient was found to decline with starvation time (postmolting time) for all life stages that molted. In most life stages, the ratio of V(CO2) to V(O2) (respiratory exchange ratio) declined over time, indicating a change in metabolic substrate with starvation. Finally, daily percent loss in body mass declined in a pattern similar to that of V(O2). The observed patterns in metabolic decline are evaluated in relation to the life history of bed bugs. In addition, the evolutionary development of these patterns is discussed. The metabolic pattern after feeding was also found to share several similarities with that of other ectothermic species.