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Secondary transmission of toxic baits in German cockroach (Dictyoptera: Blattellidae).

The death rates of three toxic gel baits (fipronil bait, 0.05%; hydramethylnon, bait 2.15%; and abamectin B1 bait, 0.05%) were estimated in Blattella germanica (L.). After ingestion, all three baits killed similar proportions of cockroaches, but they died more rapidly after ingestion of fipronil bait than after ingestion of abamectin B1 bait or of hydramethylnon bait. Laboratory experiments evaluated the impact of secondary transmission of fipronil bait and hydramethylnon bait. Mortality rates caused by secondary transmission were higher for fipronil bait than for hydramethylnon bait. Under controlled laboratory conditions, secondary transmission occurred mainly through direct contact with, or ingestion of, traces of baits dispersed in the environment by contaminated cockroaches. These traces were either deposited by "trampling" in the environment or on dead contaminated cockroaches. Social interactions and cannibalism played a minor role in secondary transmission of these two baits.

Animals↗

Comparison of two bioassay methods for determining deltamethrin resistance in German cockroaches (Blattodea: Blattellidae).

Susceptibility to deltamethrin of 10 strains of the German cockroach, Blattella germanica (L.), trapped in hotel kitchens in Singapore was determined. Two resistance detection bioassay methods (topical application and World Health Organization glass jar method) were compared. Resistance ratios based on 50% knockdown, obtained by comparison with the S strain, ranged from 17.7 to 4,235 for topical application and from 2.2 to 22 for the glass jar method. A field strain, with consistently low resistance ratios (topical method = 17.7, glass jar method = 2.2), was identified as a potential field strain to be used as a baseline for comparison with other field strains. Resistance ratios for the other field strains obtained by comparison with the R5 strain ranged from 24.5 to 239 for topical application and from 1.2 to 9.8 for the glass jar method. The results of our study demonstrate that deltamethrin-resistant German cockroaches are numerous in Singapore. Comparison between the two bioassay methods showed that there was significant correlation between KD50 and KT50 values. The glass jar method is similar to field situations but topical application is sensitive enough to define the magnitude of resistance. Differences between the two detection bioassays and the factors governing the choice of bioassay in monitoring resistance in German cockroaches are discussed.

Administration, Topical↗

Residual activity and population effects of noviflumuron for German cockroach (Dictyoptera: Blattellidae) control.

A benzoylphenyl urea insect growth regulator with the common name noviflumuron was evaluated for efficacy and residual activity on the German cockroach, Blattella germanica (L.). In laboratory studies evaluating residual activity, 0.05% noviflumuron suspension concentrate produced 100% nymphal mortality 120 d after application to steel and masonite substrates. Residual activity of noviflumuron was more variable on painted plywood substrates compared with stainless steel and masonite. In bioassay arenas, population reductions caused by noviflumuron were significantly greater than Archer and the untreated populations. After 16 wk, populations exposed to 0.05, 0.1, and 0.2% noviflumuron were reduced by 51.9 +/- 19.8, 62.2 +/- 6.5, and 62.6 +/- 18.4%, respectively. Control cockroach populations and populations exposed to 1.3% pyriproxyfen at labeled rate (Archer, 0.61 g/m2) increased by 1286.3 +/- 125.1 and 937.2 +/- 137.1%, respectively, at the end of 16 wk. A field study in multifamily housing complexes showed noviflumuron (0.2 and 0.5%) to provide 73.3 +/- 8.0 and 90.6 +/- 3.6% trap catch reduction at 4 wk posttreatment, respectively. There were no significant differences in the performance of noviflumuron, Maxforce FC Roach Bait Stations (0.05% [AI] fipronil), and Avert dust bait (0.05% [AI] abamectin B1). Noviflumuron shows excellent potential for use in cockroach management programs.

Animals↗

Effectiveness of education for control of house dust mites and cockroaches in Seoul, Korea.

We evaluated the efficacy of health education in reducing indoor arthropod allergens in Seoul. The mite control measures comprised the use of mite-proof mattress and pillow coverings, regular washing of potentially infested materials, maintenance of a low humidity, removal of carpets, and frequent vacuum cleaning. Cockroach control measures included trapping, application of insecticides, and protecting food. Of 201 homes enrolled in October 1999, 63 volunteers were included in a 2-year follow-up survey between April 2000 and January 2002. Before intervention, the density of mites/g of dust varied greatly; 27.1/g in childrenos bedding, 20/g in adult bedding, 7.2/g on the floors of childrenos bedrooms, 6.8/g in sofas, 5.9/g on the floors of adultos bedrooms, 3.9/g on living room floors, 3.7/g in carpets, and 1.9 mites/g on kitchen floors. The predominant mite species and house percentages infested were; Dermatophagoides farinae 93%, D. pteronyssinus 9%, and Tyrophagus putrescentiae 8%. Comparing 1999 and 2001 infestations, before and after 25 mo of education, mite abundance was reduced by 98%, from 23.7 to 0.57 mites/g of dust. In 1999, cockroaches were detected in 62% homes: 36% Blattella germanica and 35% Periplaneta spp., including 9% double infestations of B. germanica and P. americana. Following intervention, cockroach infestation rates decreased to 22% of houses in 2000 and 23% in 2001. We conclude that continuous and repetitive health education resulted in the effective control of domestic arthropods.

Allergens↗

Molecular characterization of American cockroach tropomyosin (Periplaneta americana allergen 7), a cross-reactive allergen.

Inhalation of allergens produced by the American cockroach (Periplaneta americana) induces IgE Ab production and the development of asthma in genetically predisposed individuals. The cloning and expression in Escherichia coli of P. americana tropomyosin allergen have been achieved. The protein shares high homology with other arthropod tropomyosins (80% identity) but less homology with vertebrate ones (50% identity). The recombinant allergen was produced in E. coli as a nonfusion protein with a yield of 9 mg/l of bacterial culture. Both natural and recombinant tropomyosins were purified by isoelectric precipitation. P. americana allergen 1 (Per a 1) and Per a 7 (tropomyosin) are to date the only cross-reacting allergens found in cockroaches. ELISA and Western blot inhibition experiments, using natural and recombinant purified tropomyosins from shrimp and cockroach, showed that tropomyosin induced cross-reactivity of IgE from patients allergic to these allergens, suggesting that this molecule could be a common allergen among invertebrates.

Allergens↗

Influence of sanitary conditions on the field performance of chlorpyrifos-based baits against American cockroaches, Periplaneta americana (L.) (Dictyoptera: Blattidae).

The role of sanitation in performance of insecticidal bait stations containing 0.5% chlorpyrifos against the American cockroach, Periplaneta americana (L.), was investigated in residential premises. Test sites were chosen from three locations in Penang Island, Malaysia, and clustered according to their sanitary conditions. Results indicated that at 1-week post-treatment, houses with good sanitary conditions showed a significantly faster reduction (P > 0.05) in the number of cockroaches trapped (> 95%) than those with moderate and poor conditions. At 6 weeks post-treatment, all houses treated with insecticidal baits showed no significant difference in terms of reduction rate of cockroach numbers (P > 0.05), irrespective of sanitary condition. However, the bait performance in houses with poor sanitary conditions could not be sustained up to 12-week post-treatment.

Animals↗

[Length polymorphism of integrated copies of R1 and R2 retrotransposons in the German cockroach (Blattella germanica) as a potential marker for population and phylogenetic studies].

Using polymerase chain reaction technique with primers flanking target sites of retrotransposons R1 and R2, integrated copies of these transposable elements were amplified in various cockroach species (Blattodea). It was shown that each species has a unique pattern of "5'-undertranscripts" with the definite set of amplified fragments of different lengths. Intraspecies polymorphism was revealed in analysis of German cockroach specimens obtained upon individual mating. This is the first report providing results of identifying, cloning, and sequencing extended fragments (5'-truncated copies) of Blatella germanica R1 and R2 retrotransposons. It may be assumed that patterns of 5'-truncated copies of R1 and R2 elements can be used as markers in population and phylogenetic studies. Moreover, cloned and sequenced fragments will be employed in our further studies for screening of the German cockroach genomic library in order to detect full-length copies in this class transposable elements.

Animals↗

Sarcocystis falcatula of opossums: transmission by cockroaches with fatal pulmonary disease in psittacine birds.

Old World psittacines experienced an acute fatal illness in outdoor breeding collections in South Florida. Toxoplasma-like organisms were found histologically in pulmonary capillaries and elsewhere. Because the organisms underwent schizogony and could not be transmitted to mice, we looked for a cause other than Toxoplasma gondii. An opossum was trapped on the premises of 1 facility and was found to be shedding sporocysts similar to Sarcocystis falcatula in its feces. Cockroaches were prevalent and suspected as transport hosts. Cockroaches that had ingested opossum feces and subsequently were fed to cockatoos induced an identical fatal illness. Obstruction of pulmonary capillaries by developing schizonts and pulmonary edema were the most important pathologic findings. The epidemic was stopped by biological insect control employing flightless chickens to reduce cockroach populations and by an electric fence restricting access of opossums to these outdoor psittacine breeding facilities.

Animals↗

Morphology and sensory modality of mushroom body extrinsic neurons in the brain of the cockroach, Periplaneta americana.

Mushroom bodies are paired centers in insect brains that are thought to be crucial in olfactory learning and memory. Early neuroanatomical descriptions suggested that the mushroom bodies comprise rather simple arrangements of nerve cells. Intrinsic neurons within each mushroom body were believed to receive olfactory afferents and to supply long, branched axons to extrinsic neurons that lead from the mushroom body into the protocerebrum. More recent suggestions that the mushroom bodies integrate several sensory modalities find support from intracellular and extracellular recordings of extrinsic neurons in the brains of crickets, honey bees, and cockroaches. Here, we describe two major classes of extrinsic neurons, simple and complex cells, in the mushroom bodies of the cockroach Periplaneta americana. Each class is defined by its pattern of branching in the brain. Simple neurons correspond to extrinsic neurons described from other species that have one set of dendrites only within the mushroom bodies. Complex extrinsic neurons possess dendrite-like branches within and outside the mushroom bodies. This arrangement may account in part for their observed multimodality, as might newly identified afferent neurons that terminate in the mushroom body lobes among the dendrites of extrinsic neurons and that respond to multimodal stimuli. Organizational complexity within the mushroom bodies is suggested by the grouping of intrinsic cell axons into discrete laminae. These are intersected by the block-like arrangements of dendritic fields of extrinsic neurons in a manner reminiscent of Purkinje cell dendrites intersecting parallel fibers in the cerebellum. The present results demonstrate that the cockroach mushroom body processes multimodal sensory information and that its neural arrangements contribute to a precise architecture consisting of discrete longitudinal and transverse subdivisions.

Animals↗

Topography of four classes of Kenyon cells in the mushroom bodies of the cockroach.

Mushroom bodies (MBs), which are higher centers in the insect brain, are implicated in associative memory and in the control of some behaviors. Intrinsic neurons of the MB, called Kenyon cells, receive synaptic inputs from axon terminals of input neurons in the calyx. Axons of Kenyon cells project into the pedunculus and to the alpha and beta lobes, where they make synaptic connections with dendrites of extrinsic (output) neurons. In this study, we examined the morphology of Kenyon cells in the cockroach by using Golgi stains and found that they can be classified into four classes (K1, K2, K3, and K4), according to the diameter, location, and morphology of the cell bodies, dendrites, and axons. The somata of Kenyon cells of different classes occupy different concentric zones; K1 cells occupy the most central zone, and K4 cells occupy the most peripheral zone. The main processes of Kenyon cells of different classes also occupy different concentric zones in the calyx. Dendrites of K2 and K3 cells are distributed throughout the calycal neuropil, whereas those of K1 and K4 cells cover the outer and inner halves of the depth of the neuropil, respectively. In the pedunculus and the alpha and beta lobes, axons of Kenyon cells of different classes occupy different zones, although the separation is not complete. A class of extrinsic neurons in the a lobe has dendrite-like arbors that cover the zones where either K1, K2, or K3 are located. These neurons probably transmit signals of each class of Kenyon cells. We conclude that, in the cockroach, four classes of Kenyon cells subdivide the cell body region, pedunculus, and lobes of the MBs, whereas subdivision is less prominent in the calycal neuropil.

Animal Structures↗

Tagma-specific distribution of FXPRLamides in the nervous system of the American cockroach.

FXPRLamide (pyrokinin) distribution in the central nervous system and major neurohaemal organs of the American cockroach and related cockroach species was investigated using immunocytochemistry and MALDI-TOF mass spectrometry. Six isoforms (Pea-PK-1 through -6) were found in different neurohaemal release sites. Pea-PK-1-4 and Pea-PK-6 are all stored in the retrocerebral complex and are all produced in cells located in both the suboesophageal ganglion (SOG) and the tritocerebrum. These pyrokinins were found to be concentrated in and around the corpora allata. No other known peptides were detectable in such high concentrations in this neurohaemal organ. They reach the corpora cardiaca/allata via the nervi corporis cardiaci-1 (NCC-1), NCC-3, and nervi corporis allati-2 (NCA-2). Abdominal perisympathetic organs contained only Pea-PK-5 and low quantities of the sequence-related Pea-PK-6. Neither Pea-PK-5 nor -PK-6 was detected in thoracic perisympathetic organs. It is likely that the expression of pyrokinins in the central nervous system is tagma (body region)-specific. Pea-PK-6 was identified during this study as follows: Ser-Glu-Ser-Glu-Val-Pro-Gly-Met-Trp-Phe-Gly-Pro-Arg-Leu-NH(2).

Amino Acid Sequence↗

On the latency and nature of phenoloxidase present in the left colleterial gland of the cockroach Periplaneta americana.

The phenoloxidase system responsible for the sclerotization of cockroach ootheca is found to be present as an inactive form in the left colleterial gland of Periplaneta americana. The supernatant fraction obtained by centrifugation of the milky white secretions contained the inactive phenoloxidase which required both sodium dodecyl sulfate (SDS) and the insoluble sediment for exhibiting enzyme activity. Bovine serum albumin could replace the sediment in the activation process. Proteins separated from the supernatant fraction by molecular sieve chromatography on Sephadex G-25 did not require either albumin or the sediment, but required SDS for exhibiting the phenoloxidase activity. Among the detergents tested, SDS (anionic) and cetylpyridinium chloride (cationic) activated the phenoloxidase, but CHAPS (zwitterionic) or nonionic detergents failed to activate the enzyme. The activation caused by SDS occurred well below the critical micellar concentration of SDS indicating that SDS is causing the activation by binding to the protein and altering its conformation. Chloroform-methanol extracts of vestibulum or right gland could replace SDS confirming the presence of endogenous activator(s) of phenoloxidase system. A variety of exogenously added lipids could activate the latent enzyme, among which linoleate, oleate, laurate, linolenate, phosphatidylethanolamine, and phosphatidylglycerol proved to be the effective activators of the latent phenoloxidase. Partially purified phenoloxidase was found to be extremely labile and lost its activity on a) freezing and thawing, b) dialysis, and c) heating for 10 min at 55 degrees C. It exhibited a pH optimum of 7 and was inhibited drastically by phenylthiourea and diethyldithiocarbamate. It readily oxidized a number of o-diphenols such as 3,4-dihydroxybenzylalcohol, 3,4-dihydroxyphenethyl alcohol, catechol, N-acetyldopamine, N-acetylnorepinephrine, dopa, dopamine, etc., but failed to oxidize both 3,4-dihydroxybenzoic acid and 3,4-dihydroxybenzaldehyde. It neither converted the typical laccase substrate syringaldazine to its quinone methide product, nor oxidized the p-diphenols, hydroquinone and methylhydroquinone. Therefore, the enzyme participating in the quinone tanning of cockroach ootheca appears to be a typical o-diphenol oxidase and not a laccase as previously thought.

Animals↗

Oxidation of 3,4-dihydroxybenzyl alcohol: a sclerotizing precursor for cockroach ootheca.

The oxidation of 3,4-dihydroxybenzyl alcohol, one of the sclerotizing precursors for the tanning of the ootheca of cockroach Periplaneta americana, is reported for the first time. Mushroom tyrosinase catalyzed oxidation of 3,4-dihydroxybenzyl alcohol generated the corresponding quinone which was found to be unstable and readily transformed to produce 3,4-dihydroxybenzaldehyde as the stable product probably through the intermediary formation of a quinone methide. Phenoloxidase isolated from the left collateral gland of P. americana also catalyzed this new reaction. When the enzymatic oxidation of 3,4-dihydroxybenzyl alcohol was performed in the presence of a test protein such as lysozyme, the reactive species formed, caused the oligomerization of test protein. Similar studies with collateral gland proteins, failed to generate oligomers, but produced insoluble polymeric proteins. The probable fate of 3,4-dihydroxybenzyl alcohol for the tanning of cockroach ootheca is discussed.

Animals↗

Optic lobe commissures in a three-dimensional brain model of the cockroach Leucophaea maderae: a search for the circadian coupling pathways.

The circadian rhythm of locomotor activity in the cockroach Leucophaea maderae is controlled by bilaterally symmetric, apparently directly coupled, circadian pacemakers in the optic lobes. Strong evidence predicts that ventromedial to the medulla, the accessory medulla with associated pigment-dispersing hormone-immunoreactive neurons is this circadian clock. In search for direct coupling pathways between both clocks, we performed horseradish peroxidase backfills from one optic stalk as well as dextran and horseradish peroxidase injections into one accessory medulla. Seven commissures with projections in the contralateral optic lobe were identified and reconstructed. Three of these commissures connected both accessory medullae. Two of these resembled the arborization pattern of the pigment-dispersing hormone-immunoreactive neurons, which are circadian pacemaker candidates in insects. This finding suggests that some of these pacemaker candidates form a direct circadian coupling pathway. For better visualization of reconstructed commissures, we implemented the reconstructions into a three-dimensional model of the cockroach brain.

Animals↗

Identification and localization of a neprilysin-like activity that degrades tachykinin-related peptides in the brain of the cockroach, Leucophaea maderae, and locust, Locusta migratoria.

Neprilysin (NEP) is an endopeptidase, which has an important role in the inactivation of mammalian tachykinins. NEP-like activity has also been found in the brain of several insects; however, the lack of information about the cellular localization of this peptidase has hindered our understanding of its role in peptidergic signaling in insects. We now provide evidence that membrane-bound NEP is involved in the inactivation of tachykinin-related peptides in the brain of the cockroach, Leucophaea maderae, and the locust, Locusta migratoria. The L. maderae enzyme cleaved the cockroach peptide LemTRP-1 and the mammalian NEP substrate [DAla(2),Leu(5)]enkephalin at the Gly-Phe peptide bond. The enzyme was acted upon by the NEP inhibitors phosphoramidon (IC(50), 0.64 microM) and thiorphan (IC(50), 1.23 microM), and the detergent-solubilized enzyme had an Mr of approximately 300,000 and a neutral pH optimum. This endopeptidase cleaved another insect tachykinin-related peptide, CavTK-II, in a predictable manner at the Ala-Phe peptide bond, suggesting that the peptidase can hydrolyse tachykinin-related peptides with different structures. NEP activity was histochemically localized in several, but not all, regions of neuropil in the brain of L. maderae, including the central body, the lobula of the optic lobe, and the tritocerebrum. All of these regions are known to receive neuronal processes containing tachykinin-related peptides. A slightly different distribution pattern for NEP was observed in the brain of L. migratoria. Again, NEP was localized to regions of the neuropil that also display tachykinin-related peptide immunoreactivity. The data reported provide evidence for an evolutionary conserved role for NEP in the inactivation of tachykinin-related peptides in the brain.

Animals↗

Three-dimensional mapping of brain neuropils in the cockroach, Diploptera punctata.

Herein, we present a complete three-dimensional (3D) map of major neuropil structures in the central brain of the cockroach Diploptera punctata. The positions of the structures have been ascertained by confocal microscopy, which, until now-for reasons of tissue opacity and nonhomogeneity-has been thought impractical in imaging fluorescently labeled structures thicker than 150 microm. In this report, however, we have used digestive enzymes and microwave-aided fixation to stain, clear, and optically section, in its entirety, an intact central brain more than 500 microm thick. The central brain from an adult female cockroach was stained thoroughly with the membrane probe NBD-ceramide and the DNA probe propidium iodide. The central brain as well as such neuropil regions as mushroom bodies, central complex, antennal glomeruli, and lobus glomerulati were individually outlined, segmented, and reconstructed in three dimensions to a spatial resolution of approximately 1 microm in the X-Y plane and 3 microm in the Z plane. The volume and surface area of each neuropil compartment were determined, and Kenyon cells of the mushroom bodies were counted. We determined that each brain hemisphere contains about 230,000 Kenyon cells, 99 antennal lobe glomeruli, and 40 lobus glomerulatus glomeruli. Segmented compartments were assigned as separate channels and merged into a single data base to reconstruct a 3D central brain containing eight different channels. This is the first 3D map at submicron resolution of an entire animal's brain that measures more than 500 microm in thickness.

Animals↗

Distribution of histamine in the cockroach brain and visual system: an immunocytochemical and biochemical study.

The distribution of histamine-immunoreactivity in the carbodiimide-fixed brain and visual system of the cockroach was revealed immunocytochemically with an antiserum against histamine (HA). Histamine levels were measured with high-pressure liquid chromatography. The results show a widespread distribution of histamine-containing somata and fibers in the brain, particularly in the visual system. The most intense immunolabeling was seen in the retinal photoreceptors and in the first optic ganglion, the lamina, where the short visual fibers make synaptic connections with the monopolar neurons, which also displayed immunofluorescence. Immunoreactive long visual fibers traversed the lamina and outer chiasma, terminating in the distal medulla. Tracts of histamine-immunopositive fibers appeared to link the optic ganglia to the protocerebrum. Prominent histamine-containing neurons were situated in the lateral protocerebrum. Immunolabeled pathways consisting of large-diameter fibers also were seen in the cockroach brain. The central parts of the brain, including the central body, were reached by thick immunoreactive fibers that gave rise to intensely fluorescent varicose processes there. In the mushroom bodies, immunoreactivity was limited to the calyces. The protocerebral bridge was nonreactive. Immunofluorescence was seen also in the antennal lobes, but not in the antennal nerves. The biochemical measurements correlated well with the immunocytochemical data. The retinas and optic lobes, measured together, contained remarkably large amounts of histamine. These results reinforce the hypothesis presented by Hardie ('87) and Elias and Evans ('83) that histamine functions as a neurotransmitter in the photoreceptors of some, if not all, insect species.

Animals↗

Morphological and physiological characterization of small multimodal ocellar interneurons in the American cockroach.

The morphology and physiology of small multimodal ocellar interneurons (SM-neurons) have been examined in the cockroach Periplaneta americana. The SM-neuron is monopolar with its cell body in the tritocerebrum near the esophagus. Its axon ascends to the protocerebrum and extends into the ocellar neuropil through the ocellar tract and the ocellar nerve. The axon sends a long collateral process towards the optic tract. The SM-neuron responded with spike discharges to various sensory stimuli. The collateral and many side branches along the axon in the deutocerebrum appear to be input regions from those sensory afferents. Cercal stimulation triggered most effectively a train of spikes in the SM-neuron: some of seven giant axons in the ventral nerve cord were involved in this pathway. Cercal stimulation also evoked depolarization in the large second order ocellar neurons (L-neurons). A possible neural connection between SM-neurons and L-neurons was examined pharmacologically. Interaction of ocellar illumination and cercal stimulation in the L-neuron was also examined. These data are discussed in relation to the ocellar function of cockroaches and in comparison with data obtained in other insects.

Animals↗