Role of horizontal gene transfer by bacteriophages in the origin of pathogenic bacteria

The review considers the involvement of bacteriophages in transferring genes, which determine bacterial pathogenicity, and the increasing role of comparative genomics and genetics of bacteria and bacteriophages in detecting new cases of horizontal gene transfer. Examples of phage participation in this process proved to a different extent are described. Emphasis is placed on the original work carried out in Russia and focused on bacteriophages (temperate transposable phages and giant virulent phi KZ-like phages) of conditional pathogen Pseudomonas aeruginosa. Consideration is given to the possible lines of further research of the role of bacteriophages in the infection process and, in particular, the role of virulent phages, whose products are similar to those of pathogenic bacteria, in modification of clinical signs of infectious diseases and in evolution. An attempt is made to predict the possible direction of pathogen evolution associated with development of new treatment strategies and generation of new specific niches.     

Pseudolysogeny of <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> bacteria infected with φKZ-like bacteriophages

In this work, a final piece of evidence proving that bacteria Pseudomonas aeruginosa are capable of transition to the pseudolysogenic state after infection with φKZ-like phages has been produced. It was shown that the decisive factor in this process is multiple infection of bacteria with bacteriophages belonging to this genus. In the course of this work, stable clinical isolates of bacteria liberating novel bacteriophages of this genus (Che2/2 and Che21/5) were detected and attributed to species φKZ and EL, respectively, according to their phenotypic characters and the results of DNA analysis. For three bacteriophages belonging to species EL (EL, RU, and Che21/5), mutants with disorders in the capability for pseudolysogenization were isolated. One of the mutants of phage EL possesses properties of virulent mutants of typical temperate phages (vir mutant). This mutant fails to form pseudolysogens and, moreover, provides the effect of dominance upon coinfection of bacteria with the wild-type phage EL, but however is unable to exhibit this effect upon joint infection of bacteria with wild-type phages of species φKZ and Lin68. It is assumed that the effect of pseudolysogeny may be connected with functioning of φKZ and EL genes that control the products similar to repressors of other phages. Because earlier wild-type φKZ-like phages were shown to be present in commercial phage-therapeutic preparations (which represents certain problems), it is expedient to use virulent mutants of phages belonging to this genus rather than phages of the wild type.     

Bacteriophage Diversity in the North Sea

In recent years interest in bacteriophages in aquatic environments has increased. Electron microscopy studies have revealed high numbers of phage particles (10⁴ to 10⁷ particles per ml) in the marine environment. However, the ecological role of these bacteriophages is still unknown, and the role of the phages in the control of bacterioplankton by lysis and the potential for gene transfer are disputed. Even the basic questions of the genetic relationships of the phages and the diversity of phage-host systems in aquatic environments have not been answered. We investigated the diversity of 22 phage-host systems after 85 phages were collected at one station near a German island, Helgoland, located in the North Sea. The relationships among the phages were determined by electron microscopy, DNA-DNA hybridization, and host range studies. On the basis of morphology, 11 phages were assigned to the virus family Myoviridae, 7 phages were assigned to the family Siphoviridae, and 4 phages were assigned to the family Podoviridae. DNA-DNA hybridization confirmed that there was no DNA homology between phages belonging to different families. We found that the 22 marine bacteriophages belonged to 13 different species. The host bacteria were differentiated by morphological and physiological tests and by 16S ribosomal DNA sequencing. All of the bacteria were gram negative, facultatively anaerobic, motile, and coccoid. The 16S rRNA sequences of the bacteria exhibited high levels of similarity (98 to 99%) with the sequences of organisms belonging to the genus Pseudoalteromonas, which belongs to the γ subdivision of the class Proteobacteria.The marine bacterial community is responsible for a considerable portion of primary production and regeneration of nutrients in the microbial loop and is associated with a great variety of marine bacteriophages (5, 12). These phages are capable of infecting a large portion of the bacterioplankton (32, 34). It is assumed that as part of the marine food web, bacteriophages play important quantitative and qualitative roles in controlling marine bacterial populations (8, 24, 34, 39, 45). The phenotypic diversity and genotypic diversity of the phage populations are related to the interaction between phages and their host organisms, which provides a tool for understanding the interaction itself (13). To estimate the influence of marine bacteriophages on the diversity of bacterioplankton, we investigated phage diversity. The virus species concept proposed by Murphy et al. (37) delineates seven different families of bacteriophages based on morphological criteria and provides criteria for new phage species based on several traits, such as DNA homologies, serological data, protein profiles, and host ranges.In this paper, we describe the diversity and genetic relationships of marine phages based on investigations of 22 representatives from 85 phage-host systems (35, 36) collected between 1988 and 1992 from waters around an island, Helgoland, located in the North Sea. All of the phages were virulent and formed plaques on their host bacteria. We assigned the phages to different virus families, species, and strains based on morphology, DNA homology, and host range. Furthermore, we characterized the phenotypic and genotypic features of the host bacteria.     

Phenogenetic Characterization of a Group of Giant φKZ-like Bacteriophages of Pseudomonas aeruginosa

A comparative study was made of a group ofPseudomonas aeruginosa virulent giant DNA bacteriophages similar to phage KZ in several genetic and phenotypic properties (particle size, particle morphology, genome size, appearance of negative colonies, high productivity, broad spectrum of lytic activity, ability to overcome the suppressing effect of plasmids, absence of several DNA restriction sites, capability of general transduction, pseudolysogeny). We have recently sequenced the phage KZ genome (288 334 bp) [J. Mol. Biol., 2002, vol. 317, pp. 1–19]. By DNA homology, the phages were assigned to three species (represented by phages KZ, Lin68, and EL, respectively) and two new genera (KZ and EL). Restriction enzyme analysis revealed the mosaic genome structure in four phages of the KZ species (KZ, Lin21, NN, and PTB80) and two phages of the EL species (EL and RU). Comparisons with respect to phage particle size, number of structural proteins, and the N-terminal sequences of the major capsid protein confirmed the phylogenetic relatedness of the phages belonging to the KZ genus. The origin and evolution of the KZ-like phages are discussed. Analysis of protein sequences encoded by the phage KZ genome made it possible to assume wide migration of the KZ-like phages (wandering phages) among various prokaryotes and possibly eukaryotes. Since the phage KZ genome codes for potentially toxic proteins, caution must be exercised in the employment of large bacteriophages in phage therapy.     

Interspecies migration and evolution of bacteriophages of the genus phiKZ: The purpose and criteria of the search for new phiKZ-like bacteriophages

Some properties of bacteriophages with large (200 kb and more) sequenced genomes have been compared. In contrast to other large bacteriophages from different families, bacteriophages active on pseudomonads of various species (phiKZ-like bacterio phages) have some common features, which suggests their phylogenetic relationship and independence of their evolution as a result of migration among bacteria of this family. Among such common features are the absence in the genomes of these phages of sites sensitive to endonuclease PstI, the absence of genes encoding DNA polymerases that are similar to the known enzymes of this type, possible dependence of replication of the phage genome on bacterial DNA polymerase, and a considerably larger average gene size as compared to that for other phages. Criteria are suggested for searching for novel phiKZ-like bacteriophages: the size of a phag e particle, production by bacteria infected with such phages of a large amount of highly viscous mucus. Taking into account the use of these bacteriophages in therapeutic preparations (due to a broad spectrum of lytic activity) and a poor knowledge of a majority of their gene products, it seems necessary to perform a more comprehensive genetic analysis of phages of this genus or their mutants for selecting those adequate for phage therapy.     

DNA homology and adsorption specificity of Pseudomonas aeruginosa virulent bacteriophages

Summary The DNA homology and adsorption specificity of newly isolated virulent bacteriophages of P. aeruginosa have been studied. On the basis of this analysis all phages were divided into four groups: k, m, mnP78-like and mnF82-like bacteriophages. DNA's of k as well as m phages were shown to possess different restriction patterns although they have an extensive homology. Unlike other groups, k phages were characterized by the presence of T4 DNA ligase-repaired, single-chain breaks.Abbreviations kbp kilobase pairs - EM electron microscopy     

Outside-host phage therapy as a biological control against environmental infectious diseases

Background

Environmentally growing pathogens present an increasing threat for human health, wildlife and food production. Treating the hosts with antibiotics or parasitic bacteriophages fail to eliminate diseases that grow also in the outside-host environment. However, bacteriophages could be utilized to suppress the pathogen population sizes in the outside-host environment in order to prevent disease outbreaks. Here, we introduce a novel epidemiological model to assess how the phage infections of the bacterial pathogens affect epidemiological dynamics of the environmentally growing pathogens. We assess whether the phage therapy in the outside-host environment could be utilized as a biological control method against these diseases. We also consider how phage-resistant competitors affect the outcome, a common problem in phage therapy. The models give predictions for the scenarios where the outside-host phage therapy will work and where it will fail to control the disease. Parameterization of the model is based on the fish columnaris disease that causes significant economic losses to aquaculture worldwide. However, the model is also suitable for other environmentally growing bacterial diseases.

Results

Transmission rates of the phage determine the success of infectious disease control, with high-transmission phage enabling the recovery of the host population that would in the absence of the phage go asymptotically extinct due to the disease. In the presence of outside-host bacterial competition between the pathogen and phage-resistant strain, the trade-off between the pathogen infectivity and the phage resistance determines phage therapy outcome from stable coexistence to local host extinction.

Conclusions

We propose that the success of phage therapy strongly depends on the underlying biology, such as the strength of trade-off between the pathogen infectivity and the phage-resistance, as well as on the rate that the phages infect the bacteria. Our results indicate that phage therapy can fail if there are phage-resistant bacteria and the trade-off between pathogen infectivity and phage resistance does not completely inhibit the pathogen infectivity. Also, the rate that the phages infect the bacteria should be sufficiently high for phage-therapy to succeed.

     

Detection of novel recombinases in bacteriophage genomes unveils Rad52, Rad51 and Gp2.5 remote homologs

Homologous recombination is a key in contributing to bacteriophages genome repair, circularization and replication. No less than six kinds of recombinase genes have been reported so far in bacteriophage genomes, two (UvsX and Gp2.5) from virulent, and four (Sak, Redβ, Erf and Sak4) from temperate phages. Using profile–profile comparisons, structure-based modelling and gene-context analyses, we provide new views on the global landscape of recombinases in 465 bacteriophages. We show that Sak, Redβ and Erf belong to a common large superfamily adopting a shortcut Rad52-like fold. Remote homologs of Sak4 are predicted to adopt a shortcut Rad51/RecA fold and are discovered widespread among phage genomes. Unexpectedly, within temperate phages, gene-context analyses also pinpointed the presence of distant Gp2.5 homologs, believed to be restricted to virulent phages. All in all, three major superfamilies of phage recombinases emerged either related to Rad52-like, Rad51-like or Gp2.5-like proteins. For two newly detected recombinases belonging to the Sak4 and Gp2.5 families, we provide experimental evidence of their recombination activity in vivo. Temperate versus virulent lifestyle together with the importance of genome mosaicism is discussed in the light of these novel recombinases. Screening for these recombinases in genomes can be performed at http://biodev.extra.cea.fr/virfam.     

Comparison of the genome for phylogenetically related bacteriophages ϕKZ and EL of <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis>: Evolutionary aspects and minimal genome size

Bacteriophages of the family Myoviridae represent one of the most widespread domains of the biosphere substantially affecting the ecological balance of microorganisms. Interestingly, sequence analysis of genomic DNAs of large bacteriophages revealed many genes coding for proteins with unknown functions. A new approach is proposed to improve the functional identification of genes. This approach is based on comparing the genome sequence for phylogenetically and morphologically related phages showing no considerable homology at the level of genomic DNA. It is assumed that gene functions essential for the development of phages of a given family are conserved and that the corresponding genes code for similar orthologous proteins even when lacking sequence homology. The genome was sequenced and compared for two Pseudomonas aeruginosa giant bacteriophages, KZ and EL, which belong to a group of KZ-related phages. A substantial difference in genome organization was observed, suggesting specific features of phage evolution. In addition, the problem of the minimal genome of the superfamily is discussed on the basis of the difference in size and structure between the KZ and EL genomes.__________Translated from Genetika, Vol. 41, No. 4, 2005, pp. 455–465.Original Russian Text Copyright © 2005 by Krylov, Pleteneva, Lavigne, Hertveldt, Volckaert, Sernova, Georgopoulos, Korchevskii, Kurochkina, Mesyanzhinov.     

Developing a bacteriophage cocktail for biocontrol of potato bacterial wilt

Bacterial wilt is a devastating disease of potato and can cause an 80% production loss. To control wilt using bacteriophage therapy, we isolated and characterized twelve lytic bacteriophages from different water sources in Kenya and China. Based on the lytic curves of the phages with the pathogen Ralstonia solanacearum, one optimal bacteriophage cocktail, P1, containing six phage isolations was formulated and used for studying wilt prevention and treatment efficiency in potato plants growing in pots. The preliminary tests showed that the phage cocktail was very effective in preventing potato bacterial wilt by injection of the phages into the plants or decontamination of sterilized soil spiked with R. solanacearum. Eighty percent of potato plants could be protected from the bacterial wilt (caused by R. solanacearum reference strain GIM1.74 and field isolates), and the P1 cocktail could kill 98% of live bacteria spiked in the sterilized soil at one week after spraying. However, the treatment efficiencies of P1 depended on the timing of application of the phages, the susceptibility of the plants to the bacterial wilt, as well as the virulence of the bacteria infected, suggesting that it is important to apply the phage therapy as soon as possible once there are early signs of the bacterial wilt. These results provide the basis for the development of bacteriophagebased biocontrol of potato bacterial wilt as an alternative to the use of antibiotics.

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Plasmid transduction in Streptococcus thermophilus

Summary Streptococcus thermophilus is a common dairy starter for which very few genetic exchange systems have been described. Here we report plasmid transduction by 17 and 56, two virulent phages of industrial yogurt starter strains. Several replicons could be transduced, independently of their size, with efficiencies which are high enough to allow gene transfer from a transformable intermediary strain of S. thermophilus to other hosts of the bacteriophages studied.     

Comparisons of the Genomes of New Giant Phages Isolated from Environmental Pseudomonas aeruginosa Strains of Different Regions

To study the genome diversity of bacteriophages from geographically distant natural populations, new giant KZ-like Pseudomonas aeruginosa phages isolated in two different regions were compared with earlier known phages of three species (KZ, Lin68, EL). A broad spectrum of lytic activity was demonstrated for all KZ-like phages. Phages of the KZ species proved to be common in natural populations of various regions, while EL- and Lin68-related phages were extremely rare. Most KZ-related phages had unique DNA restriction patterns, but the differences between these were only minor, and the genomes did not contain nonhomologous fragments. The spectrum of capsid polypeptides proved to be conserved in each species, and was proposed as a character necessary and sufficient for express classification of phages with an accuracy of species. Phages isolated in different geographical regions showed no substantial difference. Some phages only slightly differing in DNA restriction pattern from KZ may be used to study the origin of KZ genes coding for orthologs of proteins of unrelated species (other phages, pathogenic bacteria, eukaryotes).     

Bacteriophage phi297, a new species of <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> temperate phages with a mosaic genome: Potential use in phage therapy

The genome of halo-forming temperate Pseudomonas aeruginosa phage phi297 and lytic activity of its virulent mutant were studied. A mosaic structure was revealed for phi297 genome by its complete sequencing. The phi297 genome was partly homologous to the genomes of phages D3 and F116. High lytic activity was assumed for temperate P. aeruginosa bacteriophage phi297 on the basis of morphological features of negative colonies. Virulent mutant phi297vir, which was capable of lysing the wild-type phage bacteria, was isolated. Lytic activity was compared for phi297 and the phages from commercial mixtures of two manufacturers (facilities of Nizhnii Novgorod and Perm’). Phage phi297 caused lysis of the mutant PAO1 bacteria that were resistant to the phages from commercial preparations, but the lytic activity spectrum of phi297 was narrower that the spectra of the commercial phages. The use of nonreverting virulent mutants of certain temperate bacteriophages was proposed for the treatment of P. aeruginosa infections.     

Genome instability of <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> phages of the EL species: Examination of virulent mutants

The article continues a study of pseudolysogeny in Pseudominas aeruginosa infected with phiKZ-like phages of the EL species. Analysis was performed for several newly isolated vir mutants of EL phages (EL and RU) that were virulent (capable of causing lysis of bacteria infected with the wild-type phage) and a lower extent of opalescence of negative colonies (NCs). Wile-type recombinants were detected in crosses of virulent mutants of phages EL and RU to confirm the polygenic control of virulence. Since a deletion mutation was found in one of the virulent EL mutants and high genetic instability was characteristic of another mutant, a mobile genetic element was assumed to play a role in mutagenesis. Pseudolysogeny of bacteria provides for horizontal gene transfer between different bacterial strains. Hence, sequencing of the phage genome and demonstration of the lack of toxic gene products are insufficient for the phage to be included into a therapeutic mixture. To use live phages, it is essential to study in detail the possible consequences of their interaction with host bacteria.     

Study of the diversity in a group of phages of <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> species PB1 (Myoviridae) and their behavior in adsorbtion-resistant bacterial mutants

A group of 12 Pseudomonas aeruginosa virulent bacteriophages of different origin scored with regard to the plaque phenotype are assigned to PB1-like species based on the similarity in respect to morphology of particles and high DNA homology. Phages differ in restriction profile and the set of capsid major proteins. For the purpose of studying adsorption properties of these phages, 20 random spontaneous mutants of P. aeruginosa PAO1 with the disturbed adsorption placed in two groups were isolated. Mutants of the first group completely lost the ability to adsorb all phages of this species. It is assumed that their adsorption receptors are functionally inactive or lost at all, because the attempt to isolate phage mutants or detect natural phages of PB1 species capable of overcoming resistance of these bacteria failed. The second group includes five bacterial mutants resistant to the majority of phages belonging to species PB1. These mutants maintain the vigorous growth of phage SN and poor growth of phage 9/3, which forms turbid plaques with low efficiency of plating. In the background of weak growth, phage 9/3 yields plaques that grew well. The examination of the progeny of phage 9/3, which can grow on these bacteria, showed that its DNA differed from DNA of the original phage 9/3 by restriction profile and is identical to DNA of phage PB1 with regard to this trait. Data supported a suggestion that this phage variant resulted from recombination of phage 9/3 DNA with the locus of P. aeruginosa PAO1 genome encoding the bacteriocinogenic factor R. However, this variant of phage 9/3 did not manifest the ability to grow on phage-resistant mutants of the first group. Possible reasons for the difference between phages 9/3 or SN and the remaining phages of PB1 species are discussed. A preliminary formal scheme of the modular structure for adsorption receptors on the surface of P. aeruginosa PAO1 bacteria was constructed based on the analysis of growth of some other phage species on adsorption mutants of the first type.     

Sustainability of Virulence in a Phage-Bacterial Ecosystem

Virulent phages and their bacterial hosts represent an unusual sort of predator-prey system where each time a prey is eaten, hundreds of new predators are born. It is puzzling how, despite the apparent effectiveness of the phage predators, they manage to avoid driving their bacterial prey to extinction. Here we consider a phage-bacterial ecosystem on a two-dimensional (2-d) surface and show that homogeneous space in itself enhances coexistence. We analyze different behavioral mechanisms that can facilitate coexistence in a spatial environment. For example, we find that when the latent times of the phage are allowed to evolve, selection favors “mediocre killers,” since voracious phage rapidly deplete local resources and go extinct. Our model system thus emphasizes the differences between short-term proliferation and long-term ecosystem sustainability.The replication strategies of phages fall into two major categories: virulent and temperate. A temperate phage has the ability to integrate its DNA into the host chromosome, where it is then replicated along with the bacterial DNA during cell division. This strategy allows the phage to slow down or completely stop exploitation of the bacteria, thus reducing the risk of driving its host to extinction. A virulent phage lacks this ability, and it is not fully understood how they manage to coexist with their bacterial prey (4, 19). Consider, for example, the highly effective T4 phage. For the sake of argument, let us assume a burst size of 100 offspring upon lysis. On average, not more than a single phage out of each burst of 100 should survive to infect another bacterium, or else the phage would rapidly outgrow the bacteria and drive them to extinction. The half-life (t_1/2) of a free T4 phage particle has been measured to be approximately 10 days in LB at 37°C (6). Therefore, on average, at least t_1/2 × log₂(100) ≈ 2 months should pass between infections to prevent runaway phage growth—a time span that seems highly unreasonable for many of the environments where phage and bacteria interact, such as soil or biofilm. Even a more considered calculation, inserting the above half-life measurement into more realistic Lotka-Volterra-like predator-prey models (9) does not change the conclusion that T4 and other virulent phages appear to be far too effective predators for coexistence to be feasible. It is, however, an undisputed fact that virulent phages and bacteria have coexisted for eons and do so still, everywhere around us and inside us. One possible explanation for this puzzle is that bacteria constantly evolve resistance to existing phages and that the phages evolve to attack resistant bacteria in a continuous arms race. This “Red Queen” argument (23) has, however, been criticized on the grounds that the rates of evolution of phages and bacteria are not symmetric (17, 12). Recent measurements support this: in soil, phages appear to be “ahead of the bacteria in the coevolutionary arms race” (24). We therefore wish to explore mechanisms other than bacterial resistance that may promote coexistence between virulent phages and bacteria.Historically, phage-bacterial ecosystem models have ignored the issue of space, utilizing zero-dimensional approaches, such as ordinary differential equations (e.g., see references 1, 5, 13, 14, 15, and 21). However, many real phage-bacterial ecosystems are found in environments with a complex spatial structure, such as soil, biofilms, or wounds in animal and plant tissue. Schrag and Mittler (20) showed that coexistence between virulent phage and bacteria is feasible in a chemostat but not in serial cultures, due to the heterogeneous nature of the environment in the chemostat. Further, experiments done by Brockhurst et al. (3) indicate that reduced phage dispersal can prolong coexistence for virulent phage and bacteria in spatial environments by creating ephemeral refuges for the bacteria. Kerr et al. (10) introduced a simple cellular automaton to model fragmented populations of phage and bacteria in which coexistence was more easily achieved when migration was spatially restricted. Thus, the main extension to the simple predator-prey framework that we examine will be to add a spatial dimension.We construct and compare two phage-bacterial ecosystem models: one model where the phage and bacteria exist in a two-dimensional space, such as the surface of an agar gel (referred to as the “spatial model”), and the other model where the phage and bacteria are repeatedly mixed, mimicking serial cultures or a well-mixed broth (referred to as the “well-mixed model”). We show that space does indeed enhance coexistence. We then move on to explore other mechanisms that phage could incorporate into their behavior to further enhance coexistence. These can broadly be classified as “hardwired” (where every phage follows the same deterministic strategy) versus “adaptive” (where each phage potentially behaves differently, thus allowing the population to explore different options).We have chosen to look at three specific mechanisms as examples of these categories: (i) phage effectiveness would be reduced if they were unable to register whether they were infecting live, infected, or dead bacteria (a hardwired behavior); (ii) phage could prolong their latent time, concurrently increasing burst size, depending on the number of multiple infections (also a hardwired behavior, but a more “active” sort, where each phage senses and responds to information from the environment; T4 is known to use such a lysis inhibition strategy), and (iii) phage offspring could have altered latent times due to mutations in the holin genes (an adaptive behavior). We will compare each of these mechanisms in the spatial and well-mixed models to investigate whether the heterogeneity possible in a spatial environment affects the outcome.     

The isolation and characterization of two Stenotrophomonas maltophilia bacteriophages capable of cross-taxonomic order infectivity

Background

A rapid worldwide increase in the number of human infections caused by the extremely antibiotic resistant bacterium Stenotrophomonas maltophilia is prompting alarm. One potential treatment solution to the current antibiotic resistance dilemma is “phage therapy”, the clinical application of bacteriophages to selectively kill bacteria.

Results

Towards that end, phages DLP1 and DLP2 (vB_SmaS-DLP_1 and vB_SmaS-DLP_2, respectively) were isolated against S. maltophilia strain D1585. Host range analysis for each phage was conducted using 27 clinical S. maltophilia isolates and 11 Pseudomonas aeruginosa strains. Both phages exhibit unusually broad host ranges capable of infecting bacteria across taxonomic orders. Transmission electron microscopy of the phage DLP1 and DLP2 morphology reveals that they belong to the Siphoviridae family of bacteriophages. Restriction fragment length polymorphism analysis and complete genome sequencing and analysis indicates that phages DLP1 and DLP2 are closely related but different phages, sharing 96.7 % identity over 97.2 % of their genomes. These two phages are also related to P. aeruginosa phages vB_Pae-Kakheti_25 (PA25), PA73, and vB_PaeS_SCH_Ab26 (Ab26) and more distantly related to Burkholderia cepacia complex phage KL1, which together make up a taxonomic sub-family. Phages DLP1 and DLP2 exhibited significant differences in host ranges and growth kinetics.

Conclusions

The isolation and characterization of phages able to infect two completely different species of bacteria is an exciting discovery, as phages typically can only infect related bacterial species, and rarely infect bacteria across taxonomic families, let alone across taxonomic orders.

Electronic supplementary material

The online version of this article (doi:10.1186/s12864-015-1848-y) contains supplementary material, which is available to authorized users.     

Distribution of phage-resistant Pseudomonas aeruginosa strains

The sensitivity of a number of P. aeruginosa clinical strains to virulent bacteriophages has been studied. Phage-resistant strains have been found to constitute a considerable proportion among the tested P. aeruginosa strains. The strains under study fall into 19 groups differing in their sensitivity to the bacteriophages used in this investigation. The strains belonging to some groups are phenotypically identical to experimentally obtained P. aeruginosa phage-resistant mutants PAO. The use of bacteriophage mutants has made it possible to demonstrate that in most cases the resistance of P. aeruginosa natural strains to type phi k phages is due to disturbances in their adsorption, whereas their resistance to type phi m and phi mn phages is, seemingly, not linked with disturbances in their capacity for adsorption on the cell membranes of the bacteria.     

Present state of lactic acid bacteria phage taxonomy

Recent developments in molecular taxonomy of bacteriophages of lactobacilli and lactic streptococci are presented. DNA homology appears to be the most valid criterion in studying phage taxonomy. For each bacterial species, phages can be classified in a few families according to this criterion. A fair correspondence is observed between the groups differentiated by DNA-DNA hybridization and those differenciated by protein composition. In 2 bacterial species, a close relatedness between some virulent and temperate phages has been demonstrated. The presence of homologous sequences in the genome of various phages may play an important role in their evolution. For future taxonomic studies, some phages representative of DNA homology groups are proposed as reference phages.     

Isolation and characterization of lytic phages fromBacterioides ruminicola ssbrevis

Two bacteriophages (Brb01 and Brb02), lytic toBacteroides ruminicola ssbrevis AR20, were isolated from sewage water. Both phages possessed polyhedral heads and long noncontractile tails, and were classified as Siphoviridae of morphotype B1. Bacteria resistant to phages Brb01 and Brb02 arose following lysis of broth cultures. Survivors of Brb01 infection were capsulated but remained susceptible to Brb02 infection. Survivors of Brb02 infection were noncapsulated and were resistant to attack by both Brb01 and Brb02. Neither phage lysogenized the host. Both phages contained double-stranded DNA, and their restriction endonuclease digestion patterns indicated that the phage genomes were circularly permuted and terminally redundant. Phage Brb01 genome was examined in greater detail and confirmed to be circularly permuted, of size 33 kb, with a terminal redundancy of 2 kb, or 6% of the length of the genome. Circularly permuted genomes in phages of rumen bacteria do not appear to have been reported previously.At present, there is considerable interest in the genetic manipulation of rumen bacteria. The characterization of the phages described herein provides the basic information required for their use in the construction of vectors for the transfer of genetic material.