Isolation of Pseudomonas aeruginosa Specific Phages with Broad Activity Spectra

Knežević, Petar; Kostanjšek, Rok; Obreht, Dragana; Petrovic, O.

doi:10.1007/s00284-009-9417-8

Cited by 34 publications

(33 citation statements)

References 21 publications

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“…The results are in concordance with an earlier study wherein the phages of Pseudomonas were successfully isolated only from water samples, while in soil and feces no Pseudomonas specific phages were detected (22).Earlier studies attribute similar results to relatively low viable counts of phage capable of infecting specific bacteria (6), relatively low rates of phage diffusion within soil, particularly under drier conditions (18) and relatively high rates of free-phage inactivation within soil (27). There have been a number of reports of phage presence, in various environments (14,2).…”

Section: Discussionsupporting

confidence: 81%

Characterization and lytic activity of Pseudomonas fluorescens phages from sewage

Radhakrishnan¹,

Ananthasubramanian²

2012

Braz. J. Microbiol.

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Pseudomonas fluorescens phages from sewage were tested against P. fluorescens isolates of soil and sewage. The phages were characterized as to host range, morphology, structural proteins and genome fingerprint. Of the seven phages isolated, one was found to be abundant in sewage (5.9×10 7 pfu/mL), having broad host range, and distinct protein and DNA profile when compared to the other six phages. DNA restriction and protein profiles of the phages and their morphology indicate the diversity in the sewage environment. None of the isolates from the rhizosphere regions of various cultivated soils were susceptible to phages isolated from sewage.

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Section: Discussionsupporting

confidence: 81%

Characterization and lytic activity of Pseudomonas fluorescens phages from sewage

Radhakrishnan¹,

Ananthasubramanian²

2012

Braz. J. Microbiol.

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“…Although B. bronchiseptica is associated with swine, the phage isolation was unsuccessful from samples taken from swine water troughs and swine wastewater. In this regard, it must be taken into account that the samples from swine farm were filtered through a paper filter, and this pretreatment can cause a phage loss [9]. However, it is also possible that the phages were absent in the samples or that their number was very low.…”

Section: Discussionmentioning

confidence: 99%

“…After incubation, presence or absence of lysis was detected, and the obtained lytic zones were characterized as clear, clear with halo or turbid, and no reaction [9]. Each phage was tested against each bacterial strain in duplicate and in three independent experiments.…”

Section: Phage Host Rangementioning

confidence: 99%

“…Apart from the bacteriophages of human pathogens, which have been studied widely, the phages of animal pathogens and opportunistic bacteria, including B. bronchiseptica, have mainly been neglected [8][9][10]. Although B. bronchiseptica was discovered more than a century ago [11], there is no reference describing its bacteriophages, except for a few related temperate bacteriophages induced from clinical strains, belonging to family Podoviridae [12,13].…”

Section: Introductionmentioning

confidence: 99%

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The First Siphoviridae Family Bacteriophages Infecting Bordetella bronchiseptica Isolated from Environment

et al. 2016

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Bordetella bronchiseptica is a well-known etiological agent of kennel cough in dogs and cats and one of the two causative agents of atrophic rhinitis, a serious swine disease. The aim of the study was to isolate B. bronchiseptica bacteriophages from environmental samples for the first time. A total of 29 phages from 65 water samples were isolated using the strain ATCC 10580 as a host. The lytic spectra of the phages were examined at 25 and 37°C, using 12 strains of B. bronchiseptica. All phages were able to plaque on 25.0 % to 41.7 % of the strains. The selected phages showed similar morphology (Siphoviridae, morphotype B2), but variation of RFLP patterns and efficacy of plating on various strains. The partial genome sequence of phage vB_BbrS_CN1 showed its similarity to phages from genus Yuavirus. Using PCR, it was confirmed that the phages do not originate from the host strain, and environmental origin was additionally confirmed by the analysis of host genome sequence in silico and plating heated and unheated samples in parallel. Accordingly, this is the first isolation of B. bronchiseptica phages from environment and the first isolation and characterization of phages of B. bronchiseptica belonging to family Siphoviridae.

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“…Jensen et al developed a multiple-host enrichment method to identify phages capable of interclass infectivity (13), and Bielke et al used a sequential isolation method to isolate phages with intergenus infectivity (14). However, these phage isolation methods have yielded variable results, with a later experiment showing no significant difference in phage host range (15). Furthermore, multiple-host enrichments tend to dramatically reduce phage infectivity, as measured by the efficiency of plating (EOP) on nonproduction hosts (13).…”

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confidence: 99%

Isolation of Polyvalent Bacteriophages by Sequential Multiple-Host Approaches

Mathieu

et al. 2016

Appl Environ Microbiol

110

126

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Many studies on phage biology are based on isolation methods that may inadvertently select for narrow-host-range phages. Consequently, broad-host-range phages, whose ecological significance is largely unexplored, are consistently overlooked. To enhance research on such polyvalent phages, we developed two sequential multihost isolation methods and tested both culturedependent and culture-independent phage libraries for broad infectivity. Lytic phages isolated from activated sludge were capable of interspecies or even interorder infectivity without a significant reduction in the efficiency of plating (0.45 to 1.15). Two polyvalent phages (PX1 of the Podoviridae family and PEf1 of the Siphoviridae family) were characterized in terms of adsorption rate (3.54 ؋ 10 ؊10 to 8.53 ؋ 10 ؊10 ml/min), latent time (40 to 55 min), and burst size (45 to 99 PFU/cell), using different hosts. These phages were enriched with a nonpathogenic host (Pseudomonas putida F1 or Escherichia coli K-12) and subsequently used to infect model problematic bacteria. By using a multiplicity of infection of 10 in bacterial challenge tests, >60% lethality was observed for Pseudomonas aeruginosa relative to uninfected controls. The corresponding lethality for Pseudomonas syringae was ϳ50%. Overall, this work suggests that polyvalent phages may be readily isolated from the environment by using different sequential hosts, and this approach should facilitate the study of their ecological significance as well as enable novel applications.T he total bacteriophage (phage) population on Earth is estimated at 10 31 or more, making phages by far the most abundant biological entities on the planet (1, 2). As such, phages exert a significant influence over global biogeochemical cycles (3, 4) and are important drivers of bacterial diversity (5). Considering their ecological importance and value as a potential genetic resource, increasing our fundamental understanding of phage biology may facilitate the development of novel applications. However, research on phage diversity and ecology may be inadvertently limited by the use of biased isolation techniques that preferentially select for narrow-host-range phages (6, 7), while-broad hostrange phages are consistently overlooked.The classic approach to isolate and study phages is typically performed with high-density, nutrient-rich batch monocultures grown under planktonic conditions and most often results in the isolation of narrow-host-range phages (8)(9)(10).Previous studies have demonstrated the existence of polyvalent phages, among which the temperate phages P1 and Mu are the most studied (11, 12). However, their broad-host-range properties were discovered accidently, and few studies have explored methods for isolating and enriching polyvalent phages. Jensen et al. developed a multiple-host enrichment method to identify phages capable of interclass infectivity (13), and Bielke et al. used a sequential isolation method to isolate phages with intergenus infectivity (14). However, these phage isolation methods have...

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Isolation of Pseudomonas aeruginosa Specific Phages with Broad Activity Spectra

Cited by 34 publications

References 21 publications

Characterization and lytic activity of Pseudomonas fluorescens phages from sewage

Characterization and lytic activity of Pseudomonas fluorescens phages from sewage

The First Siphoviridae Family Bacteriophages Infecting Bordetella bronchiseptica Isolated from Environment

Isolation of Polyvalent Bacteriophages by Sequential Multiple-Host Approaches

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