Pathogenicity of a Highly Exopolysaccharide-producing Halomonas Strain Causing Epizootics in Larval Cultures of the Chilean Scallop Argopecten purpuratus (Lamarck, 1819).

Microb Ecol. 2008 Jun 12;

Authors: Rojas R, Miranda CD, Amaro AM

Mass mortalities of larval cultures of Chilean scallop Argopecten purpuratus have repeatedly occurred in northern Chile, characterized by larval agglutination and accumulation in the bottom of rearing tanks. The exopolysaccharide slime (EPS) producing CAM2 strain was isolated as the primary organism from moribund larvae in a pathogenic outbreak occurring in a commercial hatchery producing larvae of the Chilean scallop Argopecten purpuratus located in Bahía Inglesa, Chile. The CAM2 strain was characterized biochemically and was identified by polymerase chain reaction amplification of 16S rRNA as Halomonas sp. (Accession number DQ885389.1). Healthy 7-day-old scallop larvae cultures were experimentally infected for a 48-h period with an overnight culture of the CAM2 strain at a final concentration of ca. 10(5) cells per milliliter, and the mortality and vital condition of larvae were determined by optical and scanning electron microscopy (SEM) to describe the chronology of the disease. Pathogenic action of the CAM2 strain was clearly evidenced by SEM analysis, showing a high ability to adhere and detach larvae velum cells by using its "slimy" EPS, producing agglutination, loss of motility, and a posterior sinking of scallop larvae. After 48 h, a dense bacterial slime on the shell surface was observed, producing high percentages of larval agglutination (63.28 +/- 7.87%) and mortality (45.03 +/- 4.32%) that were significantly (P < 0.05) higher than those of the unchallenged control cultures, which exhibited only 3.20 +/- 1.40% dead larvae and no larval agglutination. Furthermore, the CAM2 strain exhibited a high ability to adhere to fiberglass pieces of tanks used for scallop larvae rearing (1.64 x 10(5) cells adhered per square millimeters at 24 h postinoculation), making it very difficult to eradicate it from the culture systems. This is the first report of a pathogenic activity on scallop larvae of Halomonas species, and it prompts the necessity of an appraisal on biofilm-producing bacteria in Chilean scallop hatcheries. PMID: 18548185 [PubMed - as supplied by publisher]

Factors Predicting Bacterial Involvement in Severe Acute Exacerbations of Chronic Obstructive Pulmonary Disease.

Respiration. 2008 Jun 12;

Authors: Nseir S, Cavestri B, di Pompeo C, Diarra M, Brisson H, Lemyze M, Roussel-Delvallez M, Durocher A

Background: Strategies aiming at reducing antibiotic use are required in the intensive care unit (ICU). Although antibiotic treatment is recommended in patients with severe exacerbation of chronic obstructive pulmonary disease (COPD), a bacterial etiology is found in only a half of these patients. Objectives: The aim of this study was to determine factors predicting bacterial isolation in severe acute exacerbations of COPD. Methods: All patients with severe acute exacerbation of COPD requiring intubation and mechanical ventilation were included in this prospective observational cohort study. At ICU admission, information on endotracheal aspirate purulence and hyperthermia was collected. In all patients, Gram stain and quantitative endotracheal aspirate culture (positive at 10(6) cfu/ml) were performed. In addition, leukocyte count, C-reactive protein and procalcitonin (PCT) levels were measured. Results: Ninety-eight severe acute exacerbations of COPD requiring intubation and mechanical ventilation were studied. Forty-nine bacteria were isolated at significant threshold in 40 exacerbations. Streptococcus pneumoniae (16%), methicillin-sensitive Staphylococcus aureus (16%) and Hemophilus influenzae (14%) were the most frequently isolated bacteria. PCT >0.5 ng/ml and positive Gram stain of endotracheal aspirate were independently associated with bacterial isolation in severe acute exacerbation of COPD. Positive Gram stain and PCT >0.5 ng/ml had a negative predictive value >95%. Similar results were found after excluding patients with prior antibiotic treatment. Conclusion: Positive Gram stain of endotracheal aspirate and PCT >0.5 ng/ml are independently associated with bacterial isolation in severe acute exacerbation of COPD. These results could be helpful for future interventional studies aiming at reducing antibiotic use in these patients. PMID: 18547944 [PubMed - as supplied by publisher]

Development of an internally controlled PCR assay for broad range detection of bacteria in platelet concentrates.

J Microbiol Methods. 2008 May 14;

Authors: Rood IG, Koppelman MH, Pettersson A, Savelkoul PH

A real-time PCR assay based on the 16S rRNA gene was optimized for the detection of a broad range of bacteria in plasma and platelet concentrates (PC). A lambda phage internal control was constructed and implemented in the assay, which made it suitable for diagnostic use. Spiking studies in plasma and PCs were performed to determine the analytical sensitivity of the assay. Thirty three colony forming units (CFU)/ml of E. coli and 72 CFU/ml of Staphylococcus epidermidis could be detected in plasma, and 97 CFU/ml of S. epidermidis in PCs. The assay detected all bacteria relevant for bacterial contamination of PCs. The short turn around time of the assay made it suitable for testing PCs for bacterial contamination prior to transfusion. PMID: 18547662 [PubMed - as supplied by publisher]

DNA Poised for Release in Bacteriophage ø29.

Structure. 2008 Jun;16(6):935-43

Authors: Tang J, Olson N, Jardine PJ, Grimes S, Anderson DL, Baker TS

We present here the first asymmetric, three-dimensional reconstruction of a tailed dsDNA virus, the mature bacteriophage varphi29, at subnanometer resolution. This structure reveals the rich detail of the asymmetric interactions and conformational dynamics of the varphi29 protein and DNA components, and provides novel insight into the mechanics of virus assembly. For example, the dodecameric head-tail connector protein undergoes significant rearrangement upon assembly into the virion. Specific interactions occur between the tightly packed dsDNA and the proteins of the head and tail. Of particular interest and novelty, an approximately 60A diameter toroid of dsDNA was observed in the connector-lower collar cavity. The extreme deformation that occurs over a small stretch of DNA is likely a consequence of the high pressure of the packaged genome. This toroid structure may help retain the DNA inside the capsid prior to its injection into the bacterial host. PMID: 18547525 [PubMed - in process]