Influence of octopamine or trehalase activity in muscle and hemolymph of the American cockroach, Periplaneta americana L.

Injection of adult male cockroaches (Periplaneta americana) with 10 μl 1 μM octopamine causes elevated activity of trehalase (α,α-trehalose glucohydrolase; EC 3.2.1.28) in hemolymph and muscle but not in gut. Tyramine, dopamine and glutamate, at the same concentration, failed to elicit any effect on trehalase activity. Determination of some kinetic parameters for muscle and hemolymph trehalase reveal that octopamine causes an increase in V_max without any significant alteration in the K_m of the enzyme for trehalose. The results are discussed in terms of the physiological significance of octopamine-mediated activation of tissue trehalase.     

Gene flow in environmental Legionella pneumophila leads to genetic and pathogenic heterogeneity within a Legionnaires’ disease outbreak

Background

Legionnaires’ disease is a severe form of pneumonia caused by the environmental bacterium Legionella pneumophila. Outbreaks commonly affect people with known risk factors, but the genetic and pathogenic complexity of L. pneumophila within an outbreak is not well understood. Here, we investigate the etiology of the major Legionnaires’ disease outbreak that occurred in Edinburgh, UK, in 2012, by examining the evolutionary history, genome content, and virulence of L. pneumophila clinical isolates.

Results

Our high resolution genomic approach reveals that the outbreak was caused by multiple genetic subtypes of L. pneumophila, the majority of which had diversified from a single progenitor through mutation, recombination, and horizontal gene transfer within an environmental reservoir prior to release. In addition, we discover that some patients were infected with multiple L. pneumophila subtypes, a finding which can affect the certainty of source attribution. Importantly, variation in the complement of type IV secretion systems encoded by different genetic subtypes correlates with virulence in a Galleria mellonella model of infection, revealing variation in pathogenic potential among the outbreak source population of L. pneumophila.

Conclusions

Taken together, our study indicates previously cryptic levels of pathogen heterogeneity within a Legionnaires’ disease outbreak, a discovery that impacts on source attribution for future outbreak investigations. Furthermore, our data suggest that in addition to host immune status, pathogen diversity may be an important influence on the clinical outcome of individual outbreak infections.

Electronic supplementary material

The online version of this article (doi:10.1186/s13059-014-0504-1) contains supplementary material, which is available to authorized users.     

Induction of hypertrehalosemia by excitation in Periplaneta americana

An excitation-induced hypertrehalosemic (EXIT) response has been demostrated in the American cockroach, Periplaneta americana. The response occurs in normal insects and in insects that were ligated at the neck 24 hr prior to experimentation. Several known and putative insect neurotransmitters were tested for their ability to elicit rapidly a hypertrehalosemic state in the cockroach. Octopamine was the most effective hypertrehalosemic agent, and produced a 100% increase in haemolymph trehalose levels within 15 min of being injected into the haemocoel. Less pronounced hypertrehalosemic responses were obtained with dopamine and tyramine, but not with synephrine or 5-hydroxytryptamine. Hypertrehalosemia was observed also with normetanephrine, but not with isoproterenol. A slight inhibition of the EXIT response occurred with the β-blocking agents, propranolol and dichloroisoproterenol, but the monoamine oxidase inhibitors, iproniazid phosphate and nialamide, were without effect. It is suggested that the EXIT-response may be one of several octopamine-mediated responses to excitation in the insect.     

Pharmacological characterization of a 5-hydroxytryptamine-sensitive receptor/adenylate cyclase complex in the mandibular closer muscles of the cricket, Gryllus domestica.

The mandibular closer muscles of the cricket, Gryllus domestica, contain a 5-hydroxytryptamine (5-HT)-sensitive receptor that is coupled to adenylate cyclase. A structure-activity study of the 5-HT molecule indicates that the integrity of the ethylamine sidegroup and the presence of a negatively charged moiety at the 5 position (-OH, -OCH3) are essential for activity. A pharmacological profile is presented for this receptor. The receptor differs from any reported mammalian 5-HT receptor in that none of the mammalian agonists tested were effective. However, the mammalian antagonists for 5-HT receptors, spiperone, mianserin, and ketanserin as well as the anti-histaminic cyproheptadine were all effective antagonists in this preparation. Preliminary analysis of antagonism, particularly by spiperone, shows that these antagonists are probably acting non-competitively. On the basis of the pharmacological data, and comparisons with other insect systems, the 5-HT receptor present in the cricket mandibular muscles has been tentatively classified as 5-HT2-like.     

The Effects of Eicosanoid Biosynthesis Inhibitors On Prophenoloxidase Activation,Phagocytosis and Cell Spreading in Galleria mellonella

The invertebrate immune system produces melanotic nodules in response to bacterial infections and this has previously been shown to be mediated by eicosanoids. Nodulation occurs in two phases: the first involves hemocyte degranulation and activation of the prophenoloxidase cascade; the second involves formation of a cellular capsule by attachment and spreading of hemocytes. We demonstrate that inhibitors of eicosanoid biosynthesis affect both of these phases of nodulation in Galleria mellonella. The phospholipase A(2) inhibitor, dexamethasone, as well as the cyclooxygenase inhibitor, indomethacin, significantly inhibit phagocytosis in vitro and prophenoloxidase activation in vivo. The inhibitory effects of dexamethasone were abolished by the addition of exogenous arachidonic acid. Furthermore, 5,8,11,14- eicosatetraynoic acid, dexamethasone and indomethacin inhibit hemocyte spreading in vitro. The findings support the idea that eicosanoid derivatives mediate both phases of the nodulation response and are consistent with previous studies which attribute roles for eicosanoids in other species as modulators of cell activity.     

The effects of sodium pentachlorophenate, diet and sampling procedure on amine and tryptophan concentrations in the brain of rainbow trout, Salmo gairdneri Richardson

The response of rainbow trout (Salmo gairdneri Richardson) to sublethal levels of sodium pentachlorophenate (NaPCP) was determined by measuring brain concentrations of the amines dopamine (DA), norepinephrine (NE), 5-hydroxytryptamine (5-HT) and the free amino acid tryptophan (TP) using high performance liquid chromatography with electrochemical detection. Sodium pentachlorophenate had no major effect on the concentrations of the biogenic amines. Mean control concentrations (ng g⁻¹ brain, S.E.) were: DA, 146.2, 9.9; NE, 205.8, 16.6; and 5-HT, 110.5, 8.8. There was a significant dose-dependent increase in TP concentration. Fish exposed to 200ng I⁻¹ NaPCP showed a 128% increase in TP level relative to the mean control concentration of 1980 (50) ng g⁻¹ brain. The effects of diets varying in lipid and carbohydrate content, anaesthetization with tricaine methanesulphonate or 2-phenoxyethanol, and electroshock were also investigated. No differences between treatment groups were detected.     

An inexpensive tag for short-term studies in milkfish (Cbanos chanos Forsskal) and in seabass (Lates calcarifer Bloch)

An opercular tag for marking adult milkfish ( Chanos chanos Forsskal) and seabass ( Lates calcarifer Bloch) is described. High tag retention and relatively low mortality rates were observed in adult fish handled two to ten times during 14-to 60-day tests. The features and advantages of the tag for marking large-sized fish in short-term studies are discussed.

Zusammenfassung

Eine preiswerte Markierung für Kurzzeitstudien des Milchfisches (Chanos chanos Forsskal) und der Centropomidae (Lates calcarifer Bloch)
Eine Kiemendeckel-Markierung für adulte Milchfische (Chanos chanos Forsskal) und Centropomidae (Lates calcacifer Bloch) wird beschrieben. Sie zeichnet sich durch gute Haltbarkeit aus und verursacht relativ geringe Mortalität bei adulten Fischen, die in Versuchen von 14 bis 60 Tage Dauer 2-bis 10mal untersucht wurden. Die Eigenschaften und Vorteile dieser Markierung für große Fische in Kurzzeitstudien werden diskutiert.

Résumé

Un marquage économique pour des études de courte durée du chanidé (Chanos chanos Forsskal) et du centropomidé (Lates calcarifer Bloch)
Un marquage d'opercule pour les chanidés (Chanos chanos Forsskal) et les centropomidés (Lates calcarifer Bloch) adultes est décrit. Une bonne conservation et une mortalité relativement basse ont été observées chez des poissons adultes examinés 2 à 10 fois pendant des expériences d'une durée de 14 à 60 jours. Les caractéristiques et les avantages du marquage de poissons de grande taille pendant des expériences d'une courte durée sont discutés.