The Susceptibility of Pseudomonas aeruginosa Strains from Cystic Fibrosis Patients to Bacteriophages

Phage therapy may become a complement to antibiotics in the treatment of chronic Pseudomonas aeruginosa infection. To design efficient therapeutic cocktails, the genetic diversity of the species and the spectrum of susceptibility to bacteriophages must be investigated. Bacterial strains showing high levels of phage resistance need to be identified in order to decipher the underlying mechanisms. Here we have selected genetically diverse P. aeruginosa strains from cystic fibrosis patients and tested their susceptibility to a large collection of phages. Based on plaque morphology and restriction profiles, six different phages were purified from “pyophage”, a commercial cocktail directed against five different bacterial species, including P. aeruginosa. Characterization of these phages by electron microscopy and sequencing of genome fragments showed that they belong to 4 different genera. Among 47 P. aeruginosa strains, 13 were not lysed by any of the isolated phages individually or by pyophage. We isolated two new phages that could lyse some of these strains, and their genomes were sequenced. The presence/absence of a CRISPR-Cas system (Clustered Regularly Interspaced Short Palindromic Repeats and Crisper associated genes) was investigated to evaluate the role of the system in phage resistance. Altogether, the results show that some P. aeruginosa strains cannot support the growth of any of the tested phages belonging to 5 different genera, and suggest that the CRISPR-Cas system is not a major defence mechanism against these lytic phages.     

Effects of marginal zinc deficiency on microtubule polymerization in the developing rat brain

One of the possible mechanisms that has been proposed to underlie the deleterious effects of zinc deficiency on brain development is an impairment in the normal formation of the cytoskeletal network. In the current study, in vivo microtubule polymerization was characterized in brain supernatant fluids, from 20-d-old pups whose dams were fed diets containing control (50 micrograms zinc/g) or marginal levels of zinc (10 micrograms zinc/g) throughout pregnancy and lactation. Pup brain and body weights were similar between the groups; however, plasma zinc concentrations were lower (27%) in pups fed the marginal zinc diet than in controls. Tubulin concentrations in 100,000 g brain supernates were similar between the groups; however, tubulin polymerization in the brain supernates was significantly lower in pups fed the marginal zinc diet compared to controls. Primarily, the early events of polymerization were affected; the lag period of the reaction was doubled, and the initial velocity was slower (26%) in supernates from pups fed the marginal zinc diet than in controls. These findings support the idea that some of the negative effects of marginal zinc deficiency on brain development and function may be mediated by an alteration in microtubule formation.     

Chemical labels differ between two closely related shearwater taxa

Chemical signals may be the basis of interspecific recognition and speciation in many animals. To test whether a chemical label allowing recognition between closely related species exists in seabirds, we investigated two closely related taxa breeding sympatrically at some localities: Cory's and Scopoli's shearwaters. Procellariiform seabirds are ideal for this study because they have a well‐developed olfactory system and unequalled associated capabilities among birds. We analysed and compared the relative volatile compounds composition of the uropygial gland secretions of Cory's and Scopoli's shearwaters. As the volatile components from uropygial secretions might also provide some critical eco‐chemical information about population origin and sex, we also examined variations in the volatile compounds between populations and sexes in Cory's shearwater. The chemical data were obtained using gas chromatography–mass spectrometry techniques looking for the presence of these three particular labels: species, population and gender. We found diagnostic differences in uropygial secretions between the two species of shearwaters and smaller but significant variation between populations of Cory's shearwater in the Atlantic. No significant differences were observed between males and females. Individuals might thus use the chemical variation between species to recognize and mate with conspecifics, especially at localities where both species breed sympatrically. Geographical variation in chemical composition of uropygial secretions suggests that selective forces might vary according to locality, and might represent a key in the species recognition. Further behavioural bioassays are needed to determine whether or not these species labels are signals allowing reproductive isolation between these two taxa. Finally, one of the aims of our study was to test easier methods than freezing for storing uropygial gland secretions in the field. We describe here a new possibility for the storage of uropygial secretion samples at ambient temperature in the field, providing an alternative, simple protocol for the sampling of avian chemosignals.     

Stalked Bacteria: Properties of Deoxriybonucleic Acid Bacteriophage φCbK

The Caulobacter crescentus bacteriophage phiCbK was studied with respect to the physical and chemical properties of both the phage and its deoxyribonucleic acid (DNA). Electron micrographs reveal the phage to be among the largest DNA bacteriophages reported, with head dimensions of 64 by 195 nm and a flexible tail 275 nm in length. The phage is composed of 57% DNA. This DNA is double-stranded as indicated by (i) the sharp increase in extinction coefficient over a narrow range of temperature increase, (ii) an increase in density in CsCl upon thermal denaturation, and (iii) the equivalence of guanine and cytosine as well as adenine and thymine, determined by chemical analysis. phiCbK DNA cosediments with Escherichia coli phage T2 DNA and has therefore been assigned an S(20,w) value of 63.5S. The size of the phage and its DNA and the percentage of DNA indicate that the phiCbK phage head is relatively loosely packed. The properties of the DNA from bacteriophage phiCbK are similar to those of host C. crescentus DNA with respect to buoyant density, thermal transition point, and guanine plus cytosine content.     

Deoxyribonucleic Acid-Dependent Ribonucleic Acid Polymerase of Caulobacter crescentus

Deoxyribonucleic acid-dependent ribonucleic acid (RNA) polymerase (EC 2.7.7.6) was purified from the dimorphic bacterium Caulobacter crescentus at three stages in development. Enzyme from pure populations of stalked cells, as well as populations enriched in swarmer and predivisional cells, appeared identical in subunit structure and template requirements. The molecular weights of the enzyme subunits were 165,000, 155,000, 101,000, and 44,000, respectively. By analogy with RNA polymerase from other bacterial sources, they are considered to be components of the C. crescentus holoenzyme, beta', beta, sigma, and alpha, respectively. The C. crescentus enzyme appeared similar to the Pseudomonas aeruginosa enzyme and unlike the Escherichia coli enzyme with respect to subunit molecular weights and failure to separate into core and sigma components upon phosphocellulose chromatography. In addition, the effects of ionic strength on the time course of polymerization varied both with the sources of bacterial polymerase and bacteriophage DNA.     

Comparative Pathogenicity of Actinomyces naeslundii and Actinomyces israelii

Typical actinomycosis has been produced in mice following single intraperitoneal injections of saline suspensions of Actinomyces israelii and A. naeslundii. A. israelii produced infections in 95.8% of the animals inoculated. A. naeslundii, generally considered to be a saprophytic organism, produced lesions in 89.7% of the inoculated animals. The finding that A. naeslundii produced lesions in mice similar to those produced by A. israelii suggests that A. naeslundii has similar pathogenic potential for man. The isolation of A. naeslundii from suppurative lesions of man also supports this conclusion.