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Rearing improvements for the German cockroach (Dictyoptera: Blattellidae).

Known-age cohorts of three strains of German cockroach, Blattella germanica (L.), were mass-reared by establishing one rearing container each week (so that cockroaches within a container would be of known age). An average of 330-443 females was placed in oviposition units within these rearing containers. Percentage hatch of oothecae was 77.49-84.63%, resulting in an average of 12,258-16,359 nymphs per rearing container. Development of nymphs was similar for the three strains. Survivorship of nymphs to the adult stage was 19.95-29.56%; survivorship was significantly higher for the strain most recently colonized from the field. Adult emergence occurred at 6-7 wk, and average production per container was 2,834-3,971 cockroaches.

Animals↗

Hemolymph transfer as an assay for immunorecognition in the cockroach Periplaneta americana and the locust Schistocerca gregaria.

A quantitative assay for measuring the number of hemocytic nodules formed in response to foreign particles and soluble molecules has been used, in the locust Schistocerca and the cockroach Periplaneta, to investigate the response to transferred hemolymph. Xenogeneic test particles, rabbit neutrophil leukocytes, stimulate formation of nodules when injected into both insect species, compared with saline-injected controls. However, the number of nodules formed in the locust in response to cockroach hemolymph is significantly reduced compared with the response to other xenogeneic cells, and it is suggested that, in view of the strong reactivity of cockroach hemocytes to locust hemolymph and plasma, a graft-versus-host response might be occurring in the recipient locust. Whole hemolymph transferred allogeneically between Periplaneta, or xenogeneically from Blatta to Periplaneta, does not stimulate a response in the recipient. This corresponds well with results from other assays for immunorecognition of transplants and is further confirmation that allogeneic and, in some combinations xenogeneic, recognition is absent in insects.

Animals↗

Contextual olfactory learning in cockroaches.

We investigated the capability of context-dependent olfactory learning in cockroaches. One group of cockroaches received training to associate peppermint odor (conditioning stimulus) with sucrose solution (appetitive unconditioned stimulus) and vanilla odor with saline solution under illumination and to associate peppermint with aversive unconditioned stimulus and vanilla with appetitive unconditioned stimulus in the dark. Another group received training with the opposite stimulus arrangement. Before training, both groups exhibited preference for vanilla over peppermint. After training, the former group preferred peppermint over vanilla under illumination but preferred vanilla over peppermint in the dark, and the latter group exhibited the opposite odor preference. We conclude that cockroaches are capable of disambiguating the meaning of conditioning stimuli according to visual context.

Animals↗

Flavobacteria as intracellular symbionts in cockroaches.

Animal cells are the sole habitat for a variety of bacteria. Molecular sequence data have been used to position a number of these intracellular microorganisms in the overall scheme of eubacterial evolution. Most of them have been classified as proteobacteria or chlamydiae. Here we present molecular evidence placing an intracellular symbiont among the flavobacteria-bacteroides. This microorganism inhabits specialized cells in the cockroach fat body and has been described as a mutualistic endosymbiont of uncertain phylogenetic position. The small subunit ribosomal DNA of these bacteria was analysed after polymerase chain reaction amplification to investigate their phylogeny. The endosymbionts of five species of cockroaches were found to make up a coherent group with no close relatives within the eubacterial phylum defined by the flavobacteria. In addition, the relationships among the endosymbionts, as revealed by DNA sequence data, appeared to be congruent with the host taxonomic relationships. Based on the host fossil record, a tentative calibration of the nucleotide substitution rate for the cockroach flavobacteria gave results congruent with those obtained for the aphid endosymbiotic proteobacteria.

Animals↗

The evolution of soil-burrowing cockroaches (Blattaria: Blaberidae) from wood-burrowing ancestors following an invasion of the latter from Asia into Australia.

Morphologically similar cockroaches in the subfamilies Panesthiinae and Geoscapheinae (Blattaria: Blaberidae) display contrasting feeding habits, behaviour and biogeographical distributions. Panesthiinae, found throughout Asia and Australia, all live in and feed on decaying wood that they burrow into. Geoscapheinae are restricted to Australia and construct and live in burrows in the soil, where they feed on dry leaves taken from the surface. A lack of knowledge about phylogenetic relationships among these cockroaches hinders an understanding of the factors that have shaped the evolution of their diverse lifestyles and biogeography. To address this issue, we sequenced three genes from representatives of nine of the 10 genera in the two subfamilies, and performed phylogenetic analyses. The well-supported topology revealed the Panesthiinae to be paraphyletic with respect to the Geoscapheinae. Soil-burrowing cockroaches appear to have evolved from a lineage of wood burrowers that invaded Australia from the north some time after the merging of the Asian and Australian tectonic plates ca. 20 Myr ago. The main factor promoting the evolution of soil burrowing is likely to have been one of the periods of strong aridity that Australia has experienced since the Miocene period.

Animals↗

Adaptive radiation within New Zealand endemic species of the cockroach genus Celatoblatta Johns (Blattidae): a response to Plio-Pleistocene mountain building and climate change.

The South Island of New Zealand offers unique opportunities to study insect evolution due to long-term physical isolation, recent alpine habitats and high levels of biotic endemism. Using DNA sequence data from cytochrome oxidase subunit 1, we investigated the phylogeographical pattern among 10 endemic cockroach species within the genus Celatoblatta Johns (Blattidae). We tested the hypothesis that an ancestral cockroach species underwent rapid speciation in response to major climatic differentiation induced by mountain building. Results suggest that speciation was a twofold process, with an interspecific radiation of Pliocene/Pleistocene age followed by intraspecific diversification during the mid Pleistocene. Average genetic distance (maximum likelihood GTR + I + Gamma) was 9.17%, with a maximum of 14.5%. Data revealed eight deep well-supported branches, each with terminal clades. Six clades were differentiated according to morphological species, while the seventh was composed of three sympatric species. We consider the latter to be a phylogenetic species, possibly as a result of hybridization within a defined geographical area. This finding seriously challenges species distinctions for these three cockroach species. Correlation between genetic distances and a Climate Similarity Index (CSI) was negative, suggesting that species found in similar habitats are also genetically closely related. A Mantel test on within-clade genetic distances vs. linear geographical distance was positive, suggesting allopatric isolation for those haplotypes. We present a model of speciation for South Island Celatoblatta.

Adaptation, Biological↗

Cockroach sensitivity in asthmatic Egyptian children.

Cockroach-specific IgE antibodies (CR-IgE) were assayed in the sera of 51 asthmatic and 33 healthy, nonallergic children. Cockroach IgE was detected in 43 asthmatic children (84%), seven of whom showed a high CR-IgE response (> or = 1.5 IU/ml). Only three of the healthy children (9%) had a positive response, and none of them were in the strongly positive category. The difference from the asthmatic group was statistically significant (P < 0.001). Children with clinically mild asthma had a significantly lower CR-IgE positivity rate than moderate and severe cases. The presence of other allergic manifestations or family history of atopy had no relationship to CR-IgE, nor did the residency, age, duration of illness, or total serum IgE levels. However, the CR-IgE titres were positively correlated with the absolute eosinophil counts. Thus, cockroach antigens are common inhalant allergens in Egyptian asthmatic children.

Air Pollutants↗

Unique accumulation of neuropeptides in an insect: FMRFamide-related peptides in the cockroach, Periplaneta americana.

FMRFamides belong to the most extensively studied neuropeptides in invertebrates and exhibit diverse physiological effects on different target organs, such as muscles, intestine and the nervous system. This study on the American cockroach confirms for the first time that extended FMRFamides occur in non-dipteran insects. By means of tandem mass spectrometry, these neuropeptides were structurally elucidated, and sequence information was used for subsequent cloning of the cockroach FMRFamide gene. This precursor gene encodes for 24 putative peptides and shows sufficient similarity with the Drosophila FMRFamide gene. Of the 24 peptides, 23 were detected by mass spectrometric methods; it is the highest number of neuropeptide forms shown to be expressed from a single precursor in any insect. The expression was traced back to single neurons in the thoracic ganglia. The unique accumulation of these FMRFamide-related peptides in thoracic perisympathetic organs provides the definite evidence for a tagma-specific distribution of peptidergic neurohormones in neurohaemal release sites of the insect CNS. Excitatory effects of the cockroach FMRFamides were observed on antenna-heart preparations. In addition, the newly described FMRFamides reduce the spike frequency of dorsal-unpaired median neurons and reduce the intracellular calcium concentration, which may affect the peripheral release of the biogenic amine octopamine.

Action Potentials↗

Selective depletion of dopamine, octopamine, and 5-hydroxytryptamine in the nervous tissue of the cockroach (Periplaneta americana).

High-performance liquid chromatography with electrochemical detection was used to measure the concentrations of 3,4-dihydroxyphenylethylamine (dopamine), 5-hydroxytryptamine (5-HT), p-hydroxyphenylethanolamine (octopamine), alpha-methyl-p-tyrosine, and tryptophan in the cerebral ganglia of cockroaches (Periplaneta americana) after peripheral administration of alpha-methyl-p-tyrosine and alpha-methyltryptophan. In addition, the levels of dopamine, 5-HT, octopamine, alpha-methyl-p-tyrosine, and tryptophan were determined after injection of alpha-methyl-p-tyrosine, 6-hydroxydopamine, or 5,7-dihydroxytryptamine directly into the cerebral ganglia by means of microinjection needles. Peripheral administration of alpha-methyl-p-tyrosine (400-1,600 micrograms/insect) caused a reduction in dopamine and 5-HT concentrations in cockroach cerebral ganglia, although the reduction in dopamine concentrations was more pronounced. Peripheral injections of alpha-methyl-p-tyrosine also reduced octopamine levels in the cerebral ganglia. Peripheral injection of alpha-methyltryptophan (400-1,600 micrograms/insect) caused a marked reduction in 5-HT and tryptophan concentrations in cockroach cerebral ganglia without altering dopamine or octopamine concentrations. Central injections of alpha-methyl-p-tyrosine (80 micrograms/insect) reduced dopamine concentrations in the cerebral ganglia. However, neither 6-hydroxydopamine (20 micrograms/insect) nor 5,7-dihydroxytryptamine (20 micrograms/insect) caused reductions in amine levels when applied near or directly into the cerebral ganglia. The results suggest that specific lesions of aminergic neurons in insects by either 6-hydroxydopamine or 5,7-dihydroxytryptamine are impractical. The specific, long-lasting depletion of 5-HT by alpha-methyltryptophan suggests that this chemical may be useful in elucidating the functions of 5-HT in insects.

5,7-Dihydroxytryptamine↗

Differential actions of fipronil and dieldrin insecticides on GABA-gated chloride channels in cockroach neurons.

Fipronil and dieldrin are known to inhibit GABA receptors in both mammals and insects. However, the mechanism of selective toxicity of these insecticides between mammals and insects remains to be seen. One possible mechanism is that insect GABA receptors are more sensitive than mammalian GABAA receptors to fipronil and dieldrin. We examined differential actions of fipronil and dieldrin on GABA-gated chloride channels in insects and compared them with the data on mammalian GABAA receptors. Neurons were acutely dissociated from the American cockroach thoracic ganglia, and currents evoked by GABA were recorded by the whole-cell patch-clamp technique. GABA-evoked currents were carried by chloride ions, blocked by picrotoxinin, but not by bicuculline. Fipronil inhibited GABA currents with an IC50 value of 28 nM, whereas dieldrin exhibited a dual action potentiation with an EC50 value of 4 nM followed by inhibition with an IC50 value of 16 nM. Fipronil and dieldrin acted on the resting receptor at comparable rates, whereas fipronil blocked the activated receptor 10 times faster than dieldrin. Fipronil inhibition was partially reversible, whereas dieldrin inhibition was irreversible. Fipronil was 59 times more potent on cockroach GABA receptors than on rat GABAA receptors. However, the potentiating and inhibitory potencies of dieldrin in cockroach GABA receptors were comparable with those in rat GABAA receptors. It was concluded that the higher toxicity of fipronil in insects than in mammals is due partially to the higher sensitivity of GABA receptors. The mechanism of dieldrin's selective toxicity must lie in factors other than the sensitivity of GABA receptors.

Animals↗

Mite, cat, and cockroach exposure, allergen sensitisation, and asthma in children: a case-control study of three schools.

BACKGROUND: The amount of allergen necessary to sensitise genetically "at risk" children is unclear. The relation between allergen exposure and asthma is also uncertain. METHODS: To ensure a wide range of allergen exposures the data from case-control studies of asthma in children aged 12-14 years attending three schools in Los Alamos, New Mexico and Central Virginia were combined. Skin prick tests to indoor and outdoor allergens and bronchial hyperreactivity to histamine were assessed in children with and without symptoms of asthma. The concentration of mite, cat, and cockroach allergens in dust from the children's homes was used as a marker of exposure. RESULTS: Three hundred and thirty two children (157 with asthmatic symptoms and 175 controls) were investigated. One hundred and eighty three were classified as atopic on the basis of allergen skin prick tests and 68 as asthmatic (symptoms plus bronchial responsiveness). The prevalence and degree of sensitisation to mite and cockroach, but not cat, was strongly associated in atopic children with increasing domestic concentrations of these allergens. Asthma was strongly associated with sensitisation to indoor allergens (p<10(-6)) and weakly to outdoor allergens (p = 0.026). There was an association between current asthma and the concentration of mite allergen amongst atopic children (p = 0.008) but not amongst those who were specifically mite sensitised (p = 0.16). CONCLUSIONS: The domestic reservoir concentration of mite and cockroach, but not cat, allergen was closely related to the prevalence of sensitisation in atopic children. However, the prevalence of current asthma had a limited relationship to these allergen measurements, suggesting that other factors play a major part in determining which allergic individuals develop asthma.

Allergens↗

Common German cockroach whole body and fecal allergens: immunoprint inhibition studies.

Previous studies have established that cockroach allergens are important sensitizing agents in the induction/exacerbation of urban asthma. The present investigation compared saline extracts of German cockroach (Blattella germanica) whole bodies (GWBE) and feces (GFE) as important sources of allergens. Both extracts were tested prior to or following gel filtration on Sephadex G-75 (Fr2). Immunoprinting of unfractionated fecal extract using 10 RAST-positive sera detected a series of allergens with pI values between 4.15-4.55 and a single allergen that focused near the cathode. Multiple allergens were detected in both GWBE-Fr2 and GFE-Fr2. Generally, most RAST-positive sera reacted similarly, although GWBE-Fr2 had more allergenic proteins than GFE-Fr2 with pI less than 4.7 and fewer allergens with pI greater than 4.7. Immunoprint inhibition of GWBE and GFE by GWBE and GFE also demonstrated allergenic similarity, although homologous extracts were more effective inhibitors than heterologous preparations. Collectively, these studies demonstrate the allergenic similarities of cockroach whole body and fecal extracts. Thus fecal material may provide an important source of sensitizing antigen.

Allergens↗

IgE antibodies reactive with silverfish, cockroach and chironomid are frequently found in mite-positive allergic patients.

Approximately 30% of the house dust mite allergic patients in The Netherlands have IgE antibodies reactive with silverfish, cockroach and/or chironomid. In allergic patients without IgE antibodies against Dermatophagoides pteronyssinus less than 5% have IgE antibodies reactive with these insects. By means of RAST inhibition studies it is shown that cross-reactivity exists between D. pteronyssinus and silverfish, cockroach or chironomid. This means that a positive RAST for silverfish, cockroach, chironomid or D. pteronyssinus cannot be taken as evidence for exposure.

Animals↗

Three-dimensional kinematics and limb kinetic energy of running cockroaches.

We tested the hypothesis that fast-running hexapeds must generate high levels of kinetic energy to cycle their limbs rapidly compared with bipeds and quadrupeds. We used high-speed video analysis to determine the three-dimensional movements of the limbs and bodies of cockroaches (Blaberus discoidalis) running on a motorized treadmill at 21 cm s-1 using an alternating tripod gait. We combined these kinematic data with morphological data to calculate the mechanical energy produced to move the limbs relative to the overall center of mass and the mechanical energy generated to rotate the body (head + thorax + abdomen) about the overall center of mass. The kinetic energy involved in moving the limbs was 8 microJ stride-1 (a power output of 21 mW kg-1, which was only approximately 13% of the external mechanical energy generated to lift and accelerate the overall center of mass at this speed. Pitch, yaw and roll rotational movements of the body were modest (less than +/- 7 degrees), and the mechanical energy required for these rotations was surprisingly small (1.7 microJ stride-1 for pitch, 0.5 microJ stride-1 for yaw and 0.4 microJ stride-1 for roll) as was the power (4.2, 1.2 and 1.1 mW kg-1, respectively). Compared at the same absolute forward speed, the mass-specific kinetic energy generated by the trotting hexaped to swing its limbs was approximately half of that predicted from data on much larger two- and four-legged animals. Compared at an equivalent speed (mid-trotting speed), limb kinetic energy was a smaller fraction of total mechanical energy for cockroaches than for large bipedal runners and hoppers and for quadrupedal trotters. Cockroaches operate at relatively high stride frequencies, but distribute ground reaction forces over a greater number of relatively small legs. The relatively small leg mass and inertia of hexapeds may allow relatively high leg cycling frequencies without exceptionally high internal mechanical energy generation.

Animals↗

Species-specific action and distribution of tachykinin-related peptides in the foregut of the cockroaches Leucophaea maderae and Periplaneta americana.

Nine tachykinin-related peptides (TRPs) have been isolated from the brain and intestine of the cockroach Leucophaea maderae. In the present investigation, two of the nine TRPs, LemTRP 1 and 5, were tested for their ability to stimulate contractions in the foregut of the cockroaches L. maderae and Periplaneta americana in vitro. The two LemTRPs and the related locust peptide locustatachykinin I (LomTK I) induced contractions in the foregut of P. americana in a dose-dependent manner, but had no myostimulatory action in L. maderae. A half-maximal response for the LemTRPs and LomTK I was obtained at 5x10(-9)mol l-1. In both species, the neuropeptide proctolin stimulated foregut contractions. Using an antiserum to LomTK I, we demonstrated that in both species there are LomTK-like-immunoreactive (LomTK-LI) cell bodies and fibers within the ganglia and nerves of the stomatogastric nervous system. However, correlated with the species-specific action of the TRPs, we found efferent LomTK-LI nerve fibers supplying muscle fibers in the foregut of P. americana, but not in L. maderae. In both cockroach species, there is a rich supply of proctolin-immunoreactive fibers to the foregut muscle. Some of the LomTK-LI fibers supplying the P. americana foregut muscle contain co-localized proctolin immunoreactivity. These fibers appear to be derived from a large cell body in the frontal ganglion which also displayed co-localized immunoreactivities. Since TRP-containing neurons are restricted to the nerves and ganglia of the stomatogastric nervous system both in P. americana and L. maderae, TRPs may be involved in the control of foregut movements in both species, but in P. americana the control may be more complex with the additional peripherally projecting LomTK-LI neurons.

Animals↗

Intestinal peptides as circulating hormones: release of tachykinin-related peptide from the locust and cockroach midgut.

Tachykinin-related peptides (TRPs) in the locust Locusta migratoria and the cockroach Leucophaea maderae have stimulatory effects on some muscles that are not innervated by TRP-containing neurons. Thus, these tissues may be affected by circulating TRPs. Here, we have investigated whether the midgut is the source of circulating TRPs. TRP-immunoreactive material in the locust midgut is found only in the endocrine cells of the gut epithelium. In both species of insect, the endocrine cells contain several isoforms of TRPs, as determined by immunocytochemistry and a combination of chromatography (HPLC) and enzyme immunoassay (ELISA). The release of TRPs was investigated by ELISA using isolated midguts of the locust and cockroach. Elevated levels of K(+) in the bathing saline induced the release of TRP from the midgut of both species. To examine the release of TRPs into the circulation in vivo, we measured haemolymph levels of TRPs in fed and starved locusts. The concentration of TRP-immunoreactive material in fed locusts was estimated to be 0.15 nmol l(-1), and this increased approximately fourfold in insects starved for 24 h. In accordance with this observation, the content of TRP-immunoreactive material in the midgut was lower in starved locusts than in fed locusts. Although part of the increased blood concentration of TRPs may be due to reduced blood volume, our data suggest that TRPs are released as hormones from the midgut of the locust and cockroach and that this release may be linked to nutritional status.

Animals↗

Evidence for a role of GABA and Mas-allatotropin in photic entrainment of the circadian clock of the cockroach Leucophaea maderae.

Accumulating evidence suggests that the accessory medulla is the location of the circadian pacemaker in the fruit fly Drosophila melanogaster and the cockroach Leucophaea maderae. gamma-Aminobutyric acid (GABA) and Mas-allatotropin are two putative neurotransmitters, in the accessory medulla in the cockroach Leucophaea maderae. Neurons immunoreactive to the neuropeptide Mas-allatotropin are local neurons with arborizations in the noduli of the accessory medulla, while GABA-immunoreactive neurons connect the noduli of the accessory medulla to the medulla and to the lamina via processes in the distal tract. Injections of GABA and Mas-allatotropin into the vicinity of the accessory medulla resulted in stable phase-dependent resetting of the circadian locomotor activity of the cockroach. The resulting phase response curves closely matched light-dependent phase response curves, suggesting that both substances play a role in circuits relaying photic information from circadian photoreceptors to the central pacemaker.

Animals↗

Levels of house dust mite-specific IgE and cockroach-specific IgE and their association with lower pulmonary function in Taiwanese children.

OBJECTIVE: Sensitization to an aeroallergen is known to diminish pulmonary function in young children and adults; however, it remains unclear whether it produces similar effects in adolescents. This study, therefore, examined the relationship between serum allergen-specific IgE levels and pulmonary function in adolescents. DESIGN: Middle-school children were invited for a physician's evaluation and pulmonary function test when not experiencing an asthma attack and for the determination of serum levels of specific IgE to common allergens. SETTING: National Cheng Kung University Hospital, Taiwan. SUBJECTS: Middle-school children in southern Taiwan, who had completed both a nationally administered Chinese version of the International Study of Asthma and Allergies in Childhood questionnaire and a pulmonary function test in October 1996. RESULTS: Forty-two then currently asthmatic children, 38 children with asthma in remission (no reported attack for > 12 months), and 69 children without asthma completed the study. Children with asthma had a significantly lower adjusted forced expiratory flow between 25% and 75% of FVC (FEF(25-75%)) and FEV(1)/FVC than children without asthma. A greater percentage of children with asthma were more sensitized to Dermatophagoides pteronyssinus (Der p), Dermatophagoides farinae (Der f), and German cockroach but not cat dander or dog dander. Children with asthma with Der f-specific IgE > 100 IU/mL, or cockroach-specific IgE > 0.7 IU/mL showed lower pulmonary function. No such association was found in children without asthma. CONCLUSION: Our findings suggest that sensitization to Der f and German cockroach was a critical factor for the lower pulmonary function observed in middle-school children with asthma.

Adolescent↗