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Behavioural observations and neurophysiological responses of cockroaches envenomated with Latrodectus katipo venom.

1. The behaviour of American cockroaches envenomated with Latrodectus katipo venom was correlated with neurophysiological events recorded from an isolated cockroach CNS preparation to which venom was applied. 2. The injection of a venom extract into cockroaches caused convulsions (3 behavioural categories: quivering, jerking and hyperextension) which eventually led to paralysis and death. 3. Changes in the spontaneous activity of nerve 5 (from the metathoracic ganglion) corresponded to the time course of envenomation behaviours. An initial increase in discharge peaked 5-10 min after application and subsequently decreased to become irreversibly blocked.

Action Potentials↗

The venom of Ampulex compressa--effects on behaviour and synaptic transmission of cockroaches.

1. The solitary wasp Ampulex compressa stings a cockroach, Periplaneta americana, twice. 2. The first sting into the ventral thorax results in a transient paralysis. During this paralysis the wasp stings the suboesophageal ganglion, which gradually results in a permanent deactivation. 3. The venom gland is a paired and highly branched organ, with a common ductus venatus. The large lumen is lined with a folded cuticula. No venom reservoir is present. 4. Extract of the venom gland induces a slow contraction of the guinea pig ileum. 5. The agonist present in the venom cannot be identified with a known agonist. 6. Venom gland extract blocks synaptic transmission from the cercal nerve to giant neurons in the sixth abdominal ganglion of the cockroach. 7. The block develops gradually, like the gradual appearance of the effects of the sting into the suboesophageal ganglion on the behaviour of the cockroach.

Animals↗

Chilling-injury and disturbance of ion homeostasis in the coxal muscle of the tropical cockroach (Nauphoeta cinerea).

Adults of warm- and cold-acclimated tropical cockroaches, Nauphoeta cinerea were exposed to low temperatures of 0 or 5 degrees C for various time intervals (hours to days). Development of chilling-injury (defects in crawling and uncoordinated movements) and mortality during the exposure were assessed and correlated with the changes in concentrations of metal ions (Na(+), K(+) and Mg(2+)) in the haemolymph and coxal muscle tissue. Warm-acclimated insects entered chill-coma at both low temperatures. In their haemolymph, the [Na(+)] and [Mg(2+)] linearly decreased and [K(+)] increased with the increasing time of exposure. The rate of concentration changes was higher at 0 than at 5 degrees C. The concentration changes resulted in gradually dissipating equilibrium potentials across the muscle cell membranes. For instance, E(K) decreased from -49.8 to -20.7 mV during 7 days at 5 degrees C. Such a disturbance of ion homeostasis was paralleled by the gradual development of chilling-injury and mortality. Most of the cockroaches showed chilling-injury when the molar ratio of [Na(+)]/[K(+)] in their haemolymph decreased from an initial of 4.4 to 2.1-2.5. In contrast, the cold-acclimated cockroaches did not enter chill-coma. They maintained constant concentrations of ions in their haemolymph, constant equilibrium potentials across muscle cell membranes and the development of chilling-injury was significantly suppressed at 5 degrees C for 7 days.

Acclimatization↗

Intracellular freezing and survival in the freeze tolerant alpine cockroach Celatoblatta quinquemaculata.

The alpine cockroach Celatoblatta quinquemaculata is common at altitudes of around 1500 m on the Rock and Pillar range of Central Otago, New Zealand where it experiences freezing conditions in the winter. The cockroach is freeze tolerant, but only to c. -9 degrees C. The cause of death at temperatures below this is unknown but likely to be due to osmotic damage to cells (shrinkage). This study compared the effect of different ice nucleation temperatures (-2 and -4 degrees C) on the viability of three types of cockroach tissue (midgut, Malpighian tubules and fat body cells) and cooling to three different temperatures (-5, -8, -12 degrees C). Two types of observations were made (i) cryomicroscope observations of ice formation and cell shrinkage (ii) cell integrity (viability) using vital stains. Cell viability decreased with lower treatment temperatures but ice nucleation temperature had no significant effect. Cryomicroscope observations showed that ice spread through tissue faster at -4 than -2 degrees C and that intracellular freezing only occurred when nucleated at -4 degrees C. From temperature records during cooling, it was observed that when freezing occurred, latent heat immediately increased the insect's body temperature close to its melting point (c. -0.3 degrees C). This "rebound" temperature was independent of nucleation temperature. Some tissues were more vulnerable to damage than others. As the gut is thought to be the site of freezing, it is significant that this tissue was the most robust. The ecological importance of the effect of nucleation temperature on survival of whole animals under field conditions is discussed.

Acclimatization↗

'Candidatus Rhabdochlamydia crassificans', an intracellular bacterial pathogen of the cockroach Blatta orientalis (Insecta: Blattodea).

The genus Rickettsiella comprises various intracellular bacterial pathogens of arthropods, exhibiting a chlamydia-like developmental cycle. Species may be divided into two main groups, the R. popilliae-R. grylli group and the R. chironomi group. Previous phylogenetic studies based on the 16S ribosomal RNA encoding gene showed that two Rickettsiella species, one from each group, belong in reality to two distantly related lineages, the gamma-Proteobacteria (R. grylli) and the Chlamydiales ('Candidatus Rhabdochlamydia porcellionis', a pathogen of terrestrial isopods). In the present work, the 16S rDNA sequence of another Rickettsiella-like species, causing abdominal swelling to its cockroach host Blatta orientalis, was determined and phylogenetic analysis performed. Identical 16S rDNA sequences of 1495 nucleotides were obtained from fat body and ovary tissues of both healthy and diseased cockroach individuals. The sequence shared only 73% of similarity with R. grylli, but 82-87% with most Chlamydiales, and even 96.3% with 'Candidatus Rhabdochlamydia porcellionis'. Phylogenetic analyses confirmed the affiliation of the cockroach pathogen within the order Chlamydiales, and based on ultrastructural characteristics and genetic analyses, we propose its inclusion in the 'Candidatus Rhabdochlamydia' as a distinct taxon, 'Candidatus Rhabdochlamydia crassificans'. These results extend our knowledge of the phylogenetic diversity of the Chlamydiales.

Animals↗

Histamine-immunoreactive neurons in the brain of the cockroach Leucophaea maderae.

Histamine is the neurotransmitter of insect photoreceptor cells but has also been found in a small number of interneurons in the insect brain. In order to investigate whether the accessory medulla (AMe), the putative circadian pacemaker of the cockroach Leucophaea maderae receives direct visual input from histaminergic photoreceptors, we analyzed the distribution of histamine-like immunoreactivity in the optic lobe and midbrain of the cockroach. Intense immunostaining was detected in photoreceptor cells of the compound eye, which terminated in the first optic neuropil, the lamina, and in a distal layer of the medulla, the second optic neuropil. Histamine immunostaining in parts of the AMe, however, originated from a centrifugal neuron of the midbrain. Within the midbrain 21-23 bilaterally symmetric pairs of cell bodies were stained. Most areas of the brain were innervated by one or more of these neurons, but the protocerebral bridge and the mushroom bodies were devoid of histamine immunoreactivity. The branching patterns of most histamine-immunoreactive neurons could be reconstructed individually. While the majority of identified neurons arborized in both brain hemispheres, five cells were local neurons of the antennal lobe. A comparison with other insect species shows striking similarities in the position of certain histamine-immunoreactive neurons, but considerable variations in the presence and branching patterns of others. The data suggest a role for histamine in a non-photic input to the circadian system of the cockroach.

Animals↗

Identification and purification of an important cross-reactive allergen from American (Periplaneta americana) and German (Blattella germanica) cockroach.

Aqueous whole body extracts from two major domiciliary cockroaches, the American, Periplaneta americana, and the German, Blattella germanica, were analyzed in crossed immunoelectrophoresis and immunoblotting. Forty-five antigens were found in P. americana and 29 in B. germanica. IgE-binding antigens were identified by crossed radioimmunoelectrophoresis with sera from 30 cockroach-allergic patients. Seven and three precipitates from P. americana and B. germanica bound significant amounts of IgE. A cross-reactive, apparently homologous allergen, from P. americana and B. germanica bound IgE from 100% and 70%, respectively, of the patients. These important allergens were tentatively named Per a I and Bla g I. The allergens were purified by sequential ion exchange, gel filtration, and isoelectric focusing. Both allergens had a molecular size of 33 to 37 kd in Sephadex G-75 gel filtration, and 28 kd in high-performance liquid chromatography gel filtration. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a minor band at approximately 25 kd, and most of the protein at 6 kd. The isoelectric point of both allergens was found be to 3.5. In amino acid analysis, the allergens were highly similar. Skin test revealed the allergens to be important in vivo sensitizing agents. The allergens may be used for environmental assays for cockroach exposure in the homes of allergic subjects.

Allergens↗

The molecular evolution of the allatostatin precursor in cockroaches.

Allatostatins (ASTs) of the Tyr/Phe-Xaa-Phe-Gly Leu/Ile-NH2 family are a group of insect neuropeptides that inhibit juvenile hormone biosynthesis by the corpora allata. We have obtained genomic DNA sequences that specify the preproallatostatin precursor for the cockroaches, Blatta orientalis, Blattella germanica, Blaberus (cranufer and Supella longipalpa. The sequences obtained are similar to those of Diploptera punctata and Periplaneta americana reported previously. The precursors of all these cockroach species are similar in size, and the organization of the ASTs that they contain (there are 13 or 14, depending on the species) have been conserved. With the sequences of these precursors, and using the homologous sequence in the orthopteran Schistocera gregari as an outgroup, a phylogenetic analysis using parsimony was carried out. The dendrograms obtained from these analyses. using the amino acid as well as the nucleotide sequences, are comparable with current models for cockroach phylogeny. Parsimony analysis was also used to study the genealogy of the different ASTs within the same precursor. Results suggest that the AST sequences were generated through a process of internal gene duplication which occurred before these species diverged from each other in evolutionary time.

Amino Acid Sequence↗

The highly conserved synapse protein SNAP-25 displays sequence variability in the cockroach Leucophaea maderae.

SNAP-25 (synaptosome-associated protein of 25 kD) is attached to the intracellular side of presynaptic membranes where it serves as a target receptor for the vesicle docking machinery prior to release of neurotransmitter. SNAP-25 displays a high degree of sequence conservation between vertebrates and Drosophila melanogaster. To obtain more information about conserved regions of SNAP-25, we have isolated cDNA clones from the cockroach Leucophaea maderae. One clone (Lm1) encoded a full-length SNAP-25 protein and its deduced amino acid sequence is 77% identical to Drosophila SNAP-25. Surprisingly, the cockroach protein is 17 amino acids shorter than Drosophila SNAP-25 at the carboxy terminus. Four other cDNA clones encode parts of SNAP-25 and each clone has distinct characteristics, including amino acid replacements and unique carboxy termini. Thus, the highly conserved protein SNAP-25 displays unexpected sequence variability in the cockroach that may indicate specialized SNAP-25 isoforms.

Amino Acid Sequence↗

Cockroach allergy.

The ubiquitous existence of cockroaches and the large-scale domestic infestation seen in inner cities make cockroach proteins a significant indoor allergen and a risk factor for asthma among inner-city residents. Studies have shown that early exposure to high levels of allergen may lead to the development of asthma in individuals with a genetic predisposition to asthma. Although field trials at cockroach abatement do not yield promising results, integrated pest management still remains the best control strategy. In highly susceptible or symptomatic patients, allergen-specific immunotherapy may be beneficial, although data are limited. As molecular techniques improve and recombinant allergens are developed, a more novel form of T-cell-specific immunotherapy may prove to be efficacious without the anaphylactic side effects seen with traditional allergy vaccines.

Allergens↗

Antagonistic effects of hypertrehalosemic neuropeptide on the activities of 6-phosphofructo-1-kinase and fructose-1,6-bisphosphatase in cockroach fat body.

Hypertrehalosemic neuropeptides from the corpora cardiaca such as the decapeptide Bld HrTH bring about a profound switch in the metabolic activity of cockroach fat body during which production of the blood sugar trehalose is stimulated while the catabolism of carbohydrate (glycolysis) is inhibited. The mechanisms of the metabolic switch are not fully understood. Incubation of isolated fat body from the cockroach Blaptica dubia with 10(-8) M Bld HrTH, for 10-60 min, stimulated glycogen breakdown and increased the content of the substrates of both the glycolytic enzyme 6-phosphofructo-1-kinase (PFK, EC 2.7.1.11) and the gluconeogenic enzyme fructose-1,6-bisphosphatase (FBPase, EC 3.1.3.11) in the tissue. The glycolytic signal fructose 2,6-bisphosphate was markedly decreased in fat body on incubation with Bld HrTH. The content of ATP was slightly reduced, while the contents of ADP and AMP were increased after incubation with the hormone. Fructose 2,6-bisphosphate is a potent activator of PFK and a strong inhibitor of FBPase purified from fat body. The activity of PFK was decreased by about 90% when the hormone-dependent changes in effectors and substrates in fat body were simulated in vitro. FBPase, in contrast, was activated about 25-fold under these conditions, suggesting the hormone to stimulate gluconeogenesis in fat body. The data support the view that fructose 2,6-bisphosphate is a pivotal intracellular messenger in the hormone-induced metabolic switch from carbohydrate degradation to trehalose production in cockroach fat body.

Adenosine Monophosphate↗

Overexpression of the cytochrome c oxidase subunit I gene associated with a pyrethroid resistant strain of German cockroaches, Blattella germanica (L.).

A cytochrome c oxidase subunit I gene (COXI) was identified and isolated as a differentially expressed gene between insecticide susceptible ACY and resistant Apyr-R German cockroach strains using PCR-selected subtractive hybridization and cDNA array techniques. The cDNA sequence of COXI has an open reading frame of 1533 nucleotides encoding a putative protein of 511 amino acid residues. Northern blot analysis indicated that levels of COXI expression were similar in three life stages (eggs, nymphs, and adults) of the susceptible ACY strain. The expression of COXI in the resistant Apyr-R strain was developmentally regulated, with low expression in eggs, an increase (approximately 1.4-fold) in nymphs, and rose to a maximum (approximately 3-fold) in both adult females and males. Comparison of COXI expression between ACY and Apyr-R strains indicated that there was no difference in the eggs of the two strains, but expression was higher (approximately 1.5-fold) in nymphs and much higher (approximately 3- to 4-fold) in adult males and females of the Apyr-R strain. The levels of COXI mRNA showed about 1.4- and 1.7-fold increase in the abdomen tissues compared with the head+thorax tissues of ACY and Apyr-R strains, respectively. Although expression patterns of COXI in head+thorax and abdomen tissues were similar (i.e. lower in the head+thorax tissues and higher in the abdomen tissues) in both the ACY and Apyr-R strains, the expression of COXI was about 2.5-fold higher in the head+thorax and approximately 3-fold higher in the abdomen tissues of the Apyr-R strain compared with the corresponding ACY samples. The overexpression of COXI in resistant German cockroaches merits the investigation of the importance of the gene in insecticide resistant German cockroaches.

Animals↗

Contamination of nebulizer equipment with cockroach allergen: there's a bug in the system!

BACKGROUND: Physicians often have anecdotal reports of patients describing increased asthma symptoms after the use of nebulizers; however, there are few published reports of nebulizer-associated exacerbations. OBJECTIVE: To present 2 cases of asthmatic children who experienced a life-threatening exacerbation of their symptoms after nebulizer use. METHODS: Case 2's nebulizer was tested for cockroach allergen by washing the medication reservoir with 2 mL of sterile filtered 1% phosphate-buffered saline, 0.05% bovine serum albumin, and Tween 20 overnight with rotation. The patient's sealed albuterol nebulizer medication was used as a control. The control albuterol and test solutions were analyzed for Blattella germanica 1 and 2 allergens using a monoclonal antibody-based immunoenzymetric assay. RESULTS: The reservoir from case 2 was found to have measurable levels of both Bla g 1 and Bla g 2. The control albuterol solution had no measurable cockroach allergen. An insect found in the nebulizer box of case 2 was identified as an infantile German cockroach. CONCLUSIONS: Nebulizer use provides an opportunity for antigen exposure directly to small airways, which may lead to severe allergic reactions in patients using contaminated equipment.

Adolescent↗

House dust mite, cat, and cockroach allergen concentrations in daycare centers in Tampa, Florida.

BACKGROUND: Allergen exposure in early childhood is a risk factor for sensitization and the development of asthma. Studies performed in Europe, New Zealand, and Singapore indicated the presence of indoor allergens in childcare centers and schools. However, the importance of indoor allergens in daycare centers in humid and warm regions of the world is not known. OBJECTIVE: To measure total mite counts, Der p 1, Der f 1, Fel d 1, and Per a 1 allergens in dust samples and mite allergen airborne concentrations in daycare centers in Tampa, Florida, United States. METHODS: Twenty daycare centers were surveyed for mite, cat, and cockroach allergens in Tampa, FL. One dust and two air samples (one during the day and one during the night) were collected in each center. Dust samples were extracted and analyzed for mite (Der p 1 and Der f 1), cat (Fel d 1), and cockroach (Per a 1) allergens. Mite airborne allergen concentrations were analyzed by RAST inhibition and expressed in standardized mite allergen units per m3 of air (AU/m3). RESULTS: Mites were identified in 15 samples, and concentrations ranged from 10 to 1,200 mites/g (298 +/- 355.2). The most prevalent mite species was Dermatophagoides pteronyssinus ( Der p 1). Der p 1 and/or Der f 1 were detected in 10 daycare centers. Der p 1 was detected in eight centers and ranged from I to 21.8 microg/g of dust (5.4 +/- 6.9); Der f 1 was detected in 3 centers and ranged from 0.2 to 2.1 microg/g of dust (1.3 +/- 0.9). Per a 1 and Fel d 1 were detected in all centers in small quantities; Per a 1 ranged from 8 to 1,806 ng/g (263.1 +/- 449.7) and Fel d 1 from 0.2 to 120 U/g of dust (16.6 +/- 31.7), respectively. Airborne mite allergen was detected in 18 centers and ranged from 0.01 to 2.7 AU/m3 during the day (0.2 +/- 0.6) and from 0.01 to 0.12 AU/m3 during the night (0.06 +/- 0.03), P = 0.001. CONCLUSIONS: Mite, cat, and cockroach allergens are present in daycare centers in Tampa, FL. Mite allergen concentrations exceeded levels that have been associated with sensitization and symptoms in allergic subjects in 40% of these centers.

Allergens↗

Microbial organisms carried by brown-banded cockroaches in relation to their spatial distribution in a hospital.

A simultaneous study of cockroach (Supella supellectilium) distribution and of associated carried bacterial flora has been made in the main hospital in Rennes (France). Wild cockroaches carry a high number of bacterial species that can be related to the normal environmental flora and a contaminant flora acquired from particular environments. The diversity of carried bacterial species reveals a proximity factor between continguous floors of the building which leads us to suppose that cockroaches are able to forage from one floor to the other.

Animals↗

Behavioral and electrophysiological evidence for volatile sex pheromones in Parcoblatta wood cockroaches.

Species within the cockroach genus Parcoblatta are sexually dimorphic for wing length; females have reduced wings and are flightless, while males have long wings that are used in flight. We predicted that Parcoblatta females would release a volatile sex pheromone to attract the more mobile males. Nymphs of the broad wood cockroach, P. lata, and the Caudell's wood cockroach, P. caudelli, were collected in forested areas in North Carolina, USA, and reared in the laboratory for observations of sexual behavior and for pheromone analysis. After several days of sexual maturation, virgin females of both species exhibited distinct calling behaviors. In females of P. lata, calling commenced 6 days after adult emergence. Under a light-dark photoperiod regime, calling behavior in both species was restricted to the scotophase. Calling consisted of a repeated pattern of raising and lowering the abdomen with occasional exposure of the genital vestibulum. To test whether calling behavior is associated with the release of pheromone, volatiles from calling and noncalling females were collected on Super-Q and tested by electroantennogram (EAG) and behavioral assays. Volatile collections from calling females elicited higher male-specific EAG responses than collections from noncalling females of the same physiological stage. In an olfactometer choice test (Y-tube), males preferred volatiles from calling females over those from noncalling females. To determine the anatomical source of the pheromone, solvent extracts of various body parts were analyzed by EAG. The first through seventh tergites were the only body parts that elicited male-specific EAG responses in both species. In P. lata, the activity of the extract increased from 1- to 7-day-old females, but was lower in mated than in virgin females of the same age. The putative pheromone gland appears to consist of numerous class-3 secretory units, each composed of a secretory cell connected to a cuticular pore via a tubular duct. We conclude that female P. lata and P. caudelli produce ex-specific volatile pheromones that are emitted during calling behavior.

Animal Communication↗

Binding characteristics of BmK I, an alpha-like scorpion neurotoxic polypeptide, on cockroach nerve cord synaptosomes.

In this study, the binding characteristics of BmK I, an alpha-like neurotoxic polypeptide purified from the venom of the Chinese scorpion Buthus martensi Karsch, were investigated on rat brain and cockroach nerve cord synaptosomes. The results showed that BmK I can bind to a single class of noninteracting binding sites on cockroach nerve cord synaptosomes with medium affinity (Kd = 16.5 +/ - 4.4 nM) and low binding capacity (Bmax = 1.05 +/- 0.23 pmol/mg protein), but lacks specific binding on rat brain synaptosomes. BmK AS, BmK AS-1 (two novel sodium channel-blocking ligands), BmK IT (an excitatory insect-selective toxin) and BmK IT2 (a depressant insect-selective toxin) from the same venom were found to be capable of depressing BmK I binding in cockroach nerve cord synaptosomes, which might be attributed to either allosteric modulation of voltage-gated Na+ channels by these toxic polypeptides or partial overlapping between the receptor binding sites of BmK I and these toxins. This thus supported the notion that alpha-like scorpion neurotoxic polypeptides bind to a distinct receptor site on sodium channels, which might be similar to the binding receptor site of alpha-type insect toxins, and also related to those of BmK AS type and insect-selective scorpion toxins on insect sodium channels.

Amino Acid Sequence↗

Exposure and allergic sensitization to cockroach allergen in East Germany.

BACKGROUND: Some studies suggest that the prevalence of sensitization to cockroach allergens may be higher in the United States than in Europe, but there are no comparable data from population-based studies. OBJECTIVES: To determine the prevalence of allergic sensitization to German cockroach (GCR) in German schoolchildren and to assess its clinical relevance; and to determine the exposure to the major GCR allergen Bla g 2 in non-selected homes and nurseries. METHODS: The prevalence of allergic sensitization to GCR and other allergens was determined by measurement of specific IgE and skin-prick tests in a cross-sectional study of 2993 children aged 5-11 years in Dresden, Germany. The prevalence of atopic disease was determined by questionnaire, and pulmonary function and bronchial hyperresponsiveness to hypertonic saline were measured. Bla g 2 exposure was determined on floors of 187 kitchens and 47 nurseries by a commercial sandwich ELISA. RESULTS: One hundred and twenty-seven (4.2%) of the children had specific IgE (> 0.7 kU/L) against GCR. Among children with current wheeze, 8.4% were GCR-sensitized. Compared to data from the United States, the prevalence of sensitization to cockroach was similar in children without asthma (3.9%), but less frequent in asthmatic children from Dresden (6.1%). After adjustment for positive reactions to other allergens (SX1 test) no significant impact of GCR sensitization on wheeze or other symptoms and diagnoses was found. Bla g 2 was detected in 29% of the kitchens and 43% of the nurseries. None of these sites had exposure levels above the proposed threshold for causing disease of 80 ng/g dust. CONCLUSION: The data suggest that allergic sensitization to GCR is less frequent in asthmatics from Dresden, Germany than in US cities. The data indicate that GCR sensitization is not an independent risk factor for asthma and other atopic diseases in 5-11-year-olds from this city.

Allergens↗