Aedes japonicus japonicus and associated woodland species attracted to Centers for Disease Control and Prevention miniature light traps baited with carbon dioxide and the Traptech mosquito lure.

Twelve reported mosquito attractants, alone or in combination, and 3 different types of traps were evaluated under field conditions for their attractiveness to host-seeking and oviposition-seeking female Aedes japonicus japonicus and associated woodland species in Windsor, CT, in 2010 and 2011. This study highlights the effectiveness of combining CO2 with the TrapTech Mosquito Lure in a Centers for Disease Control and Prevention (CDC) miniature light trap for collection of Ae. j. japonicus and associated woodland mammalian-feeding mosquitoes. The TrapTech Mosquito Lure is a proprietary blend of Bedoukian Research, Inc. It contained 250 mg of R-1-octen-3-ol and 1900 mg of ammonium bicarbonate, which were slowly released from a plastic disperser. On average, 567 Ae. j. japonicus individuals were collected per trap per night in the CDC miniature light traps baited with CO2 plus TrapTech Mosquito Lure. The numbers collected in this trap were 28 times and 100 times greater than the numbers of Ae. j. japonicus collected in the CDC miniature light trap baited only with CO2 and the gravid trap baited with hay infusion, 2 commonly used traps to assess abundance of Ae. j. japonicus. The average catches of other mammalian-biting species, Ae. cinereus, Ae. triseriatus, Ae. trivittatus, Ae. vexans, Anopheles punctipennis, An. quadrimaculatus, Coquillettidia perturbans, and Culex salinarius, were all significantly greater in the CDC miniature light trap baited with CO2 plus TrapTech Mosquito Lure than in traps with CO2 alone, but their average numbers were not as large as were those of Ae. j. japonicus. These data demonstrate that the TrapTech Mosquito Lure used in combination with CO2 in a CDC miniature light trap has potential to be a versatile and simple surveillance method for Ae. j. japonicus and other species.

Bed bug (Heteroptera: Cimicidae) attraction to pitfall traps baited with carbon dioxide, heat, and chemical lure.

Carbon dioxide (CO2), heat, and chemical lure (1-octen-3-ol and L-lactic acid) were tested as attractants for bed bugs, Cimex lectularius L. (Heteroptera: Cimicidae), by using pitfall traps. Both CO2 and heat were attractive to bed bugs. CO2 was significantly more attractive to bed bugs than heat. Traps baited with chemical lure attracted more bed bugs but at a statistically nonsignificant level. In small arena studies (56 by 44 cm), pitfall traps baited with CO2 or heat trapped 79.8 +/- 6.7 and 51.6 +/- 0.9% (mean +/- SEM) of the bed bugs after 6 h, respectively. Traps baited with CO2 + heat, CO, + chemical lure, or CO2 + heat + chemical lure captured > or = 86.7% of the bed bugs after 6 h, indicating baited pitfall traps were highly effective in attracting and capturing bed bugs from a short distance. In 3.1- by 1.8-m environmental chambers, a pitfall trap baited with CO, + heat + chemical lure trapped 57.3 +/- 6.4% of the bed bugs overnight. The pitfall trap was further tested in four bed bug-infested apartments to determine its efficacy in detecting light bed bug infestations. Visual inspections found an average of 12.0 +/- 5.4 bed bugs per apartment. The bed bugs that were found by visual inspections were hand-removed during inspections. A pitfall trap baited with CO2 and chemical lure was subsequently placed in each apartment with an average of 15.0 +/- 6.4 bed bugs collected per trap by the next morning. We conclude that baited pitfall traps are potentially effective tools for evaluating bed bug control programs and detecting early bed bug infestations.

Pseudomonas aeruginosa PqsA is an anthranilate-coenzyme A ligase.

Pseudomonas aeruginosa is an opportunistic human pathogen which relies on several intercellular signaling systems for optimum population density-dependent regulation of virulence genes. The Pseudomonas quinolone signal (PQS) is a 3-hydroxy-4-quinolone with a 2-alkyl substitution which is synthesized by the condensation of anthranilic acid with a 3-keto-fatty acid. The pqsABCDE operon has been identified as being necessary for PQS production, and the pqsA gene encodes a predicted protein with homology to acyl coenzyme A (acyl-CoA) ligases. In order to elucidate the first step of the 4-quinolone synthesis pathway in P. aeruginosa, we have characterized the function of the pqsA gene product. Extracts prepared from Escherichia coli expressing PqsA were shown to catalyze the formation of anthraniloyl-CoA from anthranilate, ATP, and CoA. The PqsA protein was purified as a recombinant His-tagged polypeptide, and this protein was shown to have anthranilate-CoA ligase activity. The enzyme was active on a variety of aromatic substrates, including benzoate and chloro and fluoro derivatives of anthranilate. Inhibition of PQS formation in vivo was observed for the chloro- and fluoroanthranilate derivatives, as well as for several analogs which were not PqsA enzymatic substrates. These results indicate that the PqsA protein is responsible for priming anthranilate for entry into the PQS biosynthetic pathway and that this enzyme may serve as a useful in vitro indicator for potential agents to disrupt quinolone signaling in P. aeruginosa.

Cloning and characterization of the NAD-dependent 7alpha-Hydroxysteroid dehydrogenase from Bacteroides fragilis.

The NAD-linked 7alpha-hydroxysteroid dehydrogenase (7-HSDH) from Bacteroides fragilis ATCC 25285 was characterized and its gene cloned. The enzyme displayed optimal activities at pH 8.5 (NAD reduction) and 6.5 (NADH oxidation). The lowest K(m) and highest V(max) values were observed with chenodeoxycholic acid and its conjugates. The protein had subunits of 27.4 kDa and a native size of 110 kDa, suggesting a homotetrameric composition. The enzyme was relatively thermostable, retaining 95% of initial activity after 1 h at 65 degrees C. A DNA probe based on the N-terminal amino acid sequence hybridized to a 2373-bp HindIII fragment of B. fragilis DNA. This fragment was cloned into E. coli and sequenced, revealing a 780-bp open reading frame. The predicted amino acid sequence of the ORF showed strong sequence similarity to three other bacterial 7-HSDHs, all in the short-chain dehydrogenase family. The regulation of expression of this gene is currently under investigation.

Functions required for extracellular quinolone signaling by Pseudomonas aeruginosa.

A set of 30 mutants exhibiting reduced production of the phenazine poison pyocyanin were isolated following transposon mutagenesis of Pseudomonas aeruginosa PAO1. The mutants could be subdivided into those with defects in the primary phenazine biosynthetic pathway and those with more pleiotropic defects. The largest set of pleiotropic mutations blocked the production of the extracellular Pseudomonas quinolone signal (PQS), a molecule required for the synthesis of secondary metabolites and extracellular enzymes. Most of these pqs mutations affected genes which appear to encode PQS biosynthetic functions, although a transcriptional regulator and an apparent response effector were also represented. Two of the genes required for PQS synthesis (phnA and phnB) had previously been assumed to encode phenazine biosynthetic functions. The transcription of one of the genes required for PQS synthesis (PA2587/pqsH) was regulated by the LasI/R quorum-sensing system, thereby linking quorum sensing and PQS regulation. Others of the pleiotropic phenazine-minus mutations appear to inactivate novel components of the quorum-sensing regulatory network, including one regulator (np20) previously shown to be required for virulence in neutropenic mice.

A bacterial cell to cell signal in the lungs of cystic fibrosis patients.

Pseudomonas aeruginosa is an opportunistic pathogen that is a major cause of mortality in cystic fibrosis (CF) patients. This bacterium has numerous genes controlled by cell to cell signaling, which occurs through a complex circuitry of interconnected regulatory systems. One of the signals is the Pseudomonas Quinolone Signal (PQS), which was identified as 2-heptyl-3-hydroxy-4-quinolone. This intercellular signal controls the expression of multiple virulence factors and is required for virulence in an insect model of P. aeruginosa infection. Previous studies have implied that the intercellular signals of P. aeruginosa are important for human disease, and our goal was to determine whether PQS was produced during human infections. In this report, three types of samples from CF patients infected with P. aeruginosa were analyzed for the presence of PQS. Sputum, bronchoalveolar lavage fluid, and mucopurulent fluid from distal airways of end-stage lungs removed at transplant, all contained PQS, indicating that this cell to cell signal is produced in vivo by P. aeruginosa infecting the lungs of CF patients.