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Salmonella in the American cockroach: evaluation of vector potential through dosed feeding experiments.

Restrained American cockroaches, Periplaneta americana L., were fed graded doses of Salmonella typhimurium ranging from 1-6 X 10(3) to 2-0 X 10(6), and their faeces assayed daily for the pathogen. Only 4 specimens out of 117 demonstrated multiplication of salmonellas, which was unrelated to size of input. When data of persistence regardless of actual numbers were expressed as percentage Salmonella-positive faecal-days, and these transformed to probits, a graph of percentage-positive faecal-days versus log dose allowed a calculation of the CD 50, or contaminative dose required for 50% of the faecal-days to be infective. The CD 50 for this cockroach species was 1-4 X 10(6) Salmonella.

Animals↗

A quantitative study of the susceptibility of cockroach species to Moniliformis moniliformis (Acanthocephala).

We exposed 19 species of cockroaches to Moniliformis moniliformis by administering shelled acanthors orally and by injecting hatched acanthors directly into the body cavity. The exposure method had little influence on susceptibility: with one exception, species that were susceptible to oral exposure were also susceptible to injected acanthors and, in no case, did acanthors develop in injected animals when they did not develop in orally exposed conspecifics. We found no support for the hypothesis that the intestine may be a barrier to M. moniliformis establishment in cockroaches. There is, however, clear interspecific variation in susceptibility to M. moniliformis infection.

Administration, Oral↗

Cockroaches as carriers of bacteria in multi-family dwellings.

The potential risk of bacterial dissemination due to the presence of cockroaches (Blattella germanica, Blattellidae) in low-income flats was investigated. Cockroaches can carry a great variety of bacterial species; we identified 30 different species from 52 different flats. Klebsiella oxycytoca, K. pneumoniae and Enterobacter cloacae were the most frequently found. Pathogenic and potentially pathogenic bacteria represented 54% of all the bacterial identifications. Bacteria were carried either on the cuticle or in the gut. Contamination through external contact is sufficient to insure bacterial diffusion. There was a very low level of overlap estimated by Pianka's index (a) between the bacterial flora of neighbouring blocks of flats, and (b) between bacterial flora of different flats in the same block.

Animals↗

Bacterial load of cockroaches in relation to urban environment.

Sanitation is an important problem in relation to the control of pests in urban environments. This investigation analysed the potential risk related to the presence of cockroaches and their capacity for disseminating bacteria in six different types of buildings: hospital nursing area and out-patient area, swimming-pool pool-side and toilet area, low-income flats and food-handling places. Fifty-six species of bacteria were identified from 157 samples, 14 of these have previously been reported as potentially pathogenic for man and vertebrates. Similarities were found between samples collected in (a) the hospital out-patient area and food-handling establishments and (b) the hospital nursing area and flats. Pool-sides possessed a poorer bacterial flora. There was a greater bacterial specific diversity in food-handling establishments, flats and swimming-bath toilet area. Enterobacter cloacae. Klebsiella pneumoniae and Klebsiella oxytoca were dominant species in flats and the hospital nursing area. Therefore, cockroaches can play a role in disseminating bacteria, which they can carry passively on their cuticle.

Animals↗

Cytotoxicity of a Quinone-containing Cockroach Sex Pheromone in Human Lung Adenocarcinoma Cells.

The cytotoxic effects of blattellaquinone (BTQ), a sex pheromone produced by adult female German cockroaches, have been studied using human lung adenocarcinoma A549 cells. 1,4-Benzoquinone (BQ), a toxic chemical implicated in benzene toxicity, was used as a reference compound. Both BQ and BTQ showed comparable toxicity toward A549 cells, with LD50 values estimated to be 14 and 19 microM, respectively. These two compounds increased the formation of an oxidized fluorescent probe, 2',7'-dichlorofluorescein, but had no effect on the cellular GSSG level. Interestingly, BTQ increased the level of 8-epi-prostaglandin F2alpha and was 4-fold more efficient in depleting cellular GSH content than BQ. Of the five GSH adducts of BTQ isolated, three were identified as mono-GSH conjugates, and the other two were di-conjugates. Mass spectrometric and NMR analyses of the di-conjugates showed that the second GSH molecule displaced the isovaleric acid moiety, potentially via a nucleophilic substitution reaction. The ability of BTQ to conjugate a second GSH molecule without quinone regeneration indicated that it may be a more effective cross-linking agent than BQ. Future experiments may be needed to evaluate the overall safety of BTQ before the commercialization of the compound as a cockroach attractant.

Adenocarcinoma↗

Oligosaccharide composition and pheromonal activity of male tergal gland secretions of the German cockroach, Blattella gennanica (L.).

In the sequential courtship behavior of the German cockroach, Blattella germanica (Dictyoptera: Blattellidae), females feed on the tergal gland secretions from the male tergites. The phagostimulative components in the male secretions were composed of a complex mixture of oligosaccharides and phospholipids. Besides seven oligosaccharides previously identified, two new trisaccharides, O-alpha-D-glucopyranosyl-(1 --> 6)-alpha-D-glucopyranosyl alpha-D-glucopyranoside and O-alpha-glucopyranosyl-(1 --> 4)-O-alpha-glucopyranosyl-(1 --> 1)-myo-inositol, were characterized as the sugar components of the secretions. The activity of the sugar components was compared in combination with the lipid fraction. Individual oligosaccharides exhibited the activity either alone or synergistically with the phospholipids. A complex mixture of these primary substances in the tergal gland secretions, thus, serves as a pheromonal phagostimulant in the precopulatory behavior, strongly eliciting feeding response in the female cockroach.

Animals↗

Comparison of oligosaccharide compositions in male nuptial secretions of three cockroach species of the genus Blattella.

Male cockroaches of the genus Blattella secrete courtship pheromones from abdominal tergal glands. Conspecific females mount courting males and feed on their tergal exudates. In the German cockroach, B. germanica, a complex mixture of oligosaccharides was previously identified as the major phagostimulants in the male tergal secretion. We compared the oligosaccharide compositions of three Blattella species from different habitats, B. germanica (house), B. lituricollis (field), and B. nipponica (forest), by gas chromatographic (GC) analysis of their trifluoroacetylated derivatives. The three possessed a series of maltooligosaccharides and oligoglucosyl trehaloses with alpha1-->4 and alpha1-->1 linkages in common. However, B. germanica was conspicuous for its accumulation of a oligoglucosyl trehalose with a alpha1-->6 linkage [O-alpha-D-glucopyranosyl-(1-->6)-alpha-D-glucopyranosyl alpha-D-glucopyranoside] as the most prominent component of the secretion. Such diversification of the male pheromonal compositions within the genus might be reflected by biosynthetic systems in males and chemosensory properties in females suited for their natural habitat.

Animals↗

Domestic allergens in public places III: house dust mite, cat, dog and cockroach allergens in British hospitals.

BACKGROUND: Exposure and sensitization to indoor allergens is a major cause of asthma. OBJECTIVES: This study investigated the levels of house dust mite, cat, dog and cockroach allergens in the dust and air in hospitals and the effects of regular vacuum cleaning on allergen levels in hospital chairs. METHODS: Der p 1, Fel d 1, Can f 1 and Bla g 2 were measured in the dust collected by vacuuming upholstered chairs and a 1 m2 area of carpet and mattress in 14 hospitals. Air samples were collected using an air sampler (flow rate 60 L/min) on 10 separate days for 4 h in the outpatient department in one of the hospitals during busy clinics when patients were waiting for their appointments. In addition, dust samples were collected on four occasions, at 4-weekly intervals, from 36 fabric covered chairs in the outpatient area of a busy chest clinic by vacuuming each chair for 2 min. During the intervening weeks, 18 of the chairs (active group) were each cleaned by vacuuming for 1 min, three times per week. Der p 1, Fel d 1, Can f 1 and Bla g 2 were assayed using monoclonal antibody-based ELISA. RESULTS: In total, 83 carpets, 69 mattresses and 42 upholstered chairs were sampled. The levels of dust mite allergen Der p 1 and cockroach allergen Bla g 2 found in the hospital setting were low. High levels of Fel d 1 (GM 22.9 microg/g, range 4.5-58) and Can f 1 (GM 21.6 microg/g, range 4-63) were found in upholstered chairs. Airborne Can f 1 was detected on every occasion (range 0.12-0.56 ng/m3), whilst detectable airborne Fel d 1 was found on 7 out of the 10 sampling days (range 0.09-0.22 ng/m3). Der p 1 and Bla g 2 were below the detection limit in all airborne samples. Following repeated vacuuming the mean cat and dog allergen levels decreased significantly (P<0.001) and were almost fivefold lower in the vacuumed chairs compared with the control group. CONCLUSIONS: Low levels of mite allergen are unlikely to be of any clinical significance to mite-sensitive asthmatic patients. However, upholstered chairs in hospitals constitute a significant reservoir of cat and dog allergen. Inhalation of airborne allergen in patients attending their hospital appointment may exacerbate asthma in those highly allergic to cats or dogs. These results question the wisdom of introducing soft furnishings and carpets into hospitals. Three-times weekly vacuuming significantly reduces allergen levels in upholstered chairs.

Air Pollution, Indoor↗

Nymphs of the Oriental cockroach (Blatta orientalis) as passive vectors of causal agents of avian tuberculosis and paratuberculosis.

The potential transmission of the causal agent of paratuberculosis Mycobacterium avium ssp. paratuberculosis and avian tuberculosis Mycobacterium avium ssp. avium (Actinomycetales: Mycobacteriaceae) by nymphs of the Oriental cockroach Blatta orientalis L. (Blattodea: Blattidae) was investigated by oral infection with mycobacterial suspensions and examination of their droppings and bodies. Both the subspecies of M. avium were isolated from droppings at 3 days post-infection and M. a. avium was found in homogenized bodies at 10 days post-infection. The identity of M. a. avium and M. a. paratuberculosis isolates was demonstrated by Restriction Fragment Length Polymorphism (RFLP) analysis. The M. a. avium isolate used as the inoculum and the isolates from the bodies and droppings of the nymphs were shown to be virulent in chickens. The results show that orally infected nymphs of B. orientalis can harbour and shed viable and virulent mycobacteria. This hazard should be considered in the implementation of control measures against mycobacterial infections of animals and humans, which should include destruction of all developmental stages of cockroaches and prevention of their access to materials that can be contaminated by mycobacteria.

Animals↗

Cockroach extract antigen increases bronchial airway epithelial permeability.

BACKGROUND: The bronchial epithelial cells of airways are subject to recurrent environmental injury throughout the life of an individual. Recently, a high incidence of asthma has been reported in inner-city children. The increased incidence of asthma in inner-city children is thought to be caused, in part, by frequent exposure to allergens of the common household pest the cockroach. OBJECTIVE: We sought to investigate whether cockroach extract antigen (CrAg) induces vascular permeability factor, also known as vascular endothelial growth factor (VEGF), and whether it increases permeability in bronchial airway epithelial cells (BAECs). METHODS: We estimated CrAg-induced VEGF release in BAECs by using an ELISA and VEGF mRNA expression by using an RT-PCR reaction. The influence of CrAg on BAEC barrier function was estimated by measuring electrical resistance with an electric cell substrate impedance-sensing system. RESULTS: Our results demonstrate that CrAg induces VEGF release in BAECs in a time-dependent manner. The VEGF induction was also confirmed by means of VEGF mRNA expression in CrAg-stimulated BAECs. CrAg decreased electrical resistance across BAEC monolayers. The maximum decrease in electrical resistance was noticed 6 hours after activation and reached a plateau thereafter. Neutralizing antibodies to VEGF significantly inhibited the decrease in BAEC electrical resistance caused by CrAg. CONCLUSIONS: These data suggest that CrAg induces VEGF release in BAECs and alters bronchial airway permeability.

Animals↗

Cytochrome P450 family 4 in a cockroach: molecular cloning and regulation by regulation by hypertrehalosemic hormone.

Hypertrehalosemic hormone (a carbohydrate-mobilizing neuroendocrine decapeptide) and starvation markedly increased levels of a cockroach (Blaberus discoidalis) fat body cytochrome P450 message. The gene represented by the cloned P450 cDNA has been named CYP4C1 (cytochrome P450 family 4, subfamily C, gene 1), a newly identified member of the ubiquitous cytochrome P450 monooxygenase gene superfamily. Blaberus CYP4C1 (511 amino acids, Mr = 58,485) has a hydrophobic NH2 terminus and a sequence near the COOH terminus that is homologous to the cysteine-containing heme-binding region definitive of cytochromes P450. The cockroach sequence is 32-36% identical to mammalian family 4A and 4B enzymes. It contains a 13-residue sequence characteristic of family 4 but not other P450s. This study suggests that CYP4C1 is hormonally regulated in association with energy substrate mobilization and supports the idea that family 4 is an old and widespread gene family.

Amino Acid Sequence↗

Biosynthesis and endocrine control of the production of the German cockroach sex pheromone 3,11-dimethylnonacosan-2-one.

The biosynthesis and endocrine regulation of sex pheromone production in the female German cockroach (Blattella germanica) were determined. Radio-TLC and radio-GLC were used to demonstrate the metabolism of 3,11-dimethylnonacosane, a major cuticular lipid component, to the corresponding alkan-2-ol and methyl ketone. [11,12-3H2]-3,11-Dimethylnonacosan-2-ol was efficiently metabolized to the methyl ketone, and radio-GLC showed that the methyl ketone product from both experiments was coeluted with a methyl ketone standard. A comparison of the metabolism of the labeled dimethylalkane and dimethylalkan-2-ol by age and sex showed that both males and females from day 1 through day 9 after adult emergence readily metabolized the alcohol to the corresponding methyl ketone, whereas only females of 5-9 days postemergence efficiently converted the labeled dimethylalkane to the corresponding methyl ketone. Application of the juvenile hormone analog hydroprene induced significant increases in the conversion of the labeled hydrocarbon to the methyl ketone in starved adult females as well as in females fed a protein-free diet, conditions under which endogenous juvenile hormone biosynthesis is nearly undetectable. These data show that the methyl ketone sex pheromone is formed by the hydroxylation and oxidation of the 3,11-dimethylalkane at the 2 position, show that the age- and sex-specific step in this process is the conversion of 3,11-dimethylnonacosane to 3,11-dimethylnonacosan-2-ol, and provide evidence that juvenile hormone regulates sex pheromone production in the German cockroach.

Animals↗

Callatostatins: neuropeptides from the blowfly Calliphora vomitoria with sequence homology to cockroach allatostatins.

Five neuropeptides with C-terminal amino acid sequence homology to cockroach allatostatins have been identified in the blowfly Calliphora vomitoria. Three have the same pentapeptide C-terminal amino acid sequence as allatostatin 1 of the cockroach Diploptera punctata. A hexadecapeptide designated callatostatin 1, isolated from thoracic ganglia, brains, and heads, has the sequence Asp-Pro-Leu-Asn-Glu-Glu-Arg-Arg-Ala-Asn-Arg-Tyr-Gly-Phe-Gly-Leu-NH2. Callatostatins 2 and 3 have been isolated from heads and thoracic ganglia, respectively; they comprise the last 14 and 8 residues of callatostatin 1. Callatostatin 4, isolated from thoracic ganglia, has the sequence Xaa-Arg-Pro-Tyr-Ser-Phe-Gly-Leu-NH2, where Xaa is either Asp or Asn. This peptide, with a serine substitution for glycine at position 5, has a C-terminal pentapeptide sequence identical to that of allatostatins 3 and 4 of D. punctata. Callatostatin 5, with the sequence Gly-Pro-Pro-Tyr-Asp-Phe-Gly-Met-NH2, was identified from whole flies. All five peptides inhibit juvenile hormone production by the corpora allata of D. punctata in vitro. Callatostatin 5 was the most potent allatostatin so far tested in this species, with maximum inhibition occurring at 1 nM. In contrast, none of the callatostatins or the allatostatins showed allatostatic activity in mature female C. vomitoria when tested at concentrations of 100 to 0.1 microM. In accordance with these results, immunoreactivity to an antiserum directed against the common C terminus of callatostatin 1 and allatostatin 1 was observed in the corpora allata of D. punctata but not in the corpus allatum of C. vomitoria, despite its presence in neurons of the brain. Neurons in the thoracic ganglion of C. vomitoria that are immunoreactive against this antiserum project to the hindgut, rectum, rectal papillae, and oviduct, suggestive of a function different from that of a true allatostatin.

Amino Acid Sequence↗

Expression of the allatostatin gene in endocrine cells of the cockroach midgut.

Cockroach allatostatins are neuropeptides that have been isolated from the brain of Diploptera punctata and shown to inhibit juvenile hormone production by the corpora allata. Enzyme-linked immunoassay and immunocytochemistry with antisera to two allatostatins, ASB2 (AYSYVSEYKRLPVYNFGL-NH2) and ASAL (APSGAQRLYGFGL-NH2), revealed that allatostatins were located not only in the insect brain but also in several peripheral tissues including the cockroach midgut and hindgut. Allatostatin-like immunoreactivity was found in nerve fibers of the stomatogastric nervous system as well as in intrinsic endocrine cells of the midgut. Midgut extracts were shown to be biologically active in an allatostatin bioassay and to contain several allatostatin-like peptides, including the octadecapeptide ASB2, which was identified by mass spectrometry following HPLC purification. Reverse transcription of brain mRNA followed by PCR with degenerate oligonucleotides for ASB2 and ASAL yielded a 338-bp fragment of the allatostatin gene that encoded six allatostatins. In situ hybridization with this probe confirmed that an allatostatin gene is expressed in intrinsic endocrine cells of the midgut. Reverse transcription of midgut mRNA followed by PCR and sequencing of the product revealed that the same gene is expressed in the midgut and in the brain. Allatostatins are thus an example of insect "brain-gut peptides" and we suggest that their function may not be restricted to the regulation of juvenile hormone production.

Amino Acid Sequence↗

Neural organization of the circadian system of the cockroach Leucophaea maderae.

The cockroach Leucophaea maderae was the first animal in which lesion experiments localized an endogenous circadian clock to a particular brain area, the optic lobe. The neural organization of the circadian system, however, including entrainment pathways, coupling elements of the bilaterally distributed internal clock, and output pathways controlling circadian locomotor rhythms are only recently beginning to be elucidated. As in flies and other insect species, pigment-dispersing hormone (PDH)-immunoreactive neurons of the accessory medulla of the cockroach are crucial elements of the circadian system. Lesions and transplantation experiments showed that the endogeneous circadian clock of the brain resides in neurons associated with the accessory medulla. The accessory medulla is organized into a nodular core receiving photic input, and into internodular and peripheral neuropil involved in efferent output and coupling input. Photic entrainment of the clock through compound eye photoreceptors appears to occur via parallel, indirect pathways through the medulla. Light-like phase shifts in circadian locomotor activity after injections of gamma-aminobutyric acid (GABA)- or Mas-allatotropin into the vicinity of the accessory medulla suggest that both substances are involved in photic entrainment Extraocular, cryptochrome-based photoreceptors appear to be present in the optic lobe, but their role in photic entrainment has not been examined. Pigment-dispersing hormone-immunoreactive neurons provide efferent output from the accessory medulla to several brain areas and to the peripheral visual system. Pigment-dispersing hormone-immunoreactive neurons, and additional heterolateral neurons are, furthermore, involved in bilateral coupling of the two pacemakers. The neuronal organization, as well as the prominent involvement of GABA and neuropeptides, shows striking similarities to the organization of the suprachiasmatic nucleus, the circadian clock of the mammalian brain.

Animals↗

Bystander T cells participate in specific response to cockroach antigen (CR) in vitro.

Allergic reactions due to whole body, body parts and fecal products of cockroach (CR) are characterized by inflammatory reaction that may lead to symptoms of rhinitis or asthma in atopic individuals. Although the majority of T cells at the site of CR hypersensitivity are not antigen specific, the cellular subset and cytokine receptors that participate and control the outcome of the reaction have not been fully studied. In this study, we have used fluorescent activated cell sorter (FACS) analysis to characterize the activation marker and cytokine profile of antigen specific and bystander T cells after in vitro stimulation of peripheral blood lymphocytes with whole body extract of CR antigen. There was significant enhancement of CD69 on blast and bystander T cells in all atopic subjects compared to non-atopics. Both antigen specific and bystander T cells showed increased expression of HLA-DR, CD25 and CD71 in 9 of 11 atopic patients compared to control. There was also an increase in CD45RA+ and a decrease in CD45RO+ cells following antigen stimulation. These results correlated with the increase in the early apoptotic cells observed in patients as measured by Annexin V stain. Our data revealed that there was no difference in the expression of CD95 in both stimulated and bystander T cells. However, there was enhancement of FasL by CR antigen, suggesting that the increased apoptosis that was observed was probably due to the Fas/FasL interaction. Positive intracellular IL2, IL-4 and IFN-gamma in T cells were observed in only the antigen specific blast cells in 83% of patients studied. These results suggest interplay of memory T cell response, apoptosis, and activated bystander T cells activities in maintaining cellular homeostasis during allergic reaction in cockroach sensitive atopic subjects.

Adolescent↗

Dynamics of cockroach ocellar neurons.

The incremental responses from the second-order neurons of the ocellus of the cockroach, Periplaneta americana, have been measured. The stimulus was a white-noise-modulated light with various mean illuminances. The kernels, obtained by cross-correlating the white-noise input against the resulting response, provided a measure of incremental sensitivity as well as of response dynamics. We found that the incremental sensitivity of the second-order neurons was an exact Weber-Fechner function; white-noise-evoked responses from second-order neurons were linear; the dynamics of second-order neurons remain unchanged over a mean illuminance range of 4 log units; the small nonlinearity in the response of the second-order neuron was a simple amplitude compression; and the correlation between the white-noise input and spike discharges of the second-order neurons produced a first-order kernel similar to that of the cell's slow potential. We conclude that signal processing in the cockroach ocellus is simple but different from that in other visual systems, including vertebrate retinas and insect compound eyes, in which the system's dynamics depend on the mean illuminance.

Acoustic Stimulation↗

Dynamic relationship between the slow potential and spikes in cockroach ocellar neurons.

The relationship between the slow potential and spikes of second-order ocellar neurons of the cockroach, Periplaneta americana, was studied. The stimulus was a sinusoidally modulated light with various mean illuminances. A solitary spike was generated at the depolarizing phase of the modulation response. Analysis of the relationship between the amplitude/frequency of voltage modulation and the rate of spike generation showed that (a) the spike initiation process was bandpass at approximately 0.5-5 Hz, (b) the process contained a dynamic linearity and a static nonlinearity, and (c) the spike threshold at optimal frequencies (0.5-5 Hz) remained unchanged over a mean illuminance range of 3.6 log units, whereas (d) the spike threshold at frequencies of less than 0.5 Hz was lower at a dimmer mean illuminance. The voltage noise in the response was larger and the mean membrane potential level was more positive at a dimmer mean illuminance. Steady or noise current injection during sinusoidal light stimulation showed that (a) the decrease in the spike threshold at a dimmer mean illuminance was due to the increase in the noise variance: the noise had facilitatory effects on the spike initiation; and (b) the change in the mean potential level had little effect on the spike threshold. We conclude that fundamental signal modifications occur during the spike initiation in the cockroach ocellar neuron, a finding that differs from the spike initiation process in other visual systems, including Limulus eye and vertebrate retina, in which it is presumed that little signal modification occurs at the analog-to-digital conversion process.

Animals↗