Bdellovibrio Strains Research Articles

Using autochthonous Bdellovibrio as a predatory bacterium for reduction of Gram-negative pathogenic bacteria in urban wastewater and reuse it.

Background and Objectives:The microbial contamination of wastewater is associated with health risks. The aim of this study was to use the autochthonous Bdellovibrio potential to prey Gram-negative pathogenic bacteria as a bio-control agent to treat urban wastewater.Materials and Methods:Thirty-six raw sewage samples were collected for isolation of Bdellovibrio. Double layer plaque assay was used for isolation and the isolates were identified by microscopic examination and molecular analysis. To evaluate the predatory potential for decrease number of Gram-negative pathogenic bacteria, plaque perdition assay, reduction in host cells viability by colony-forming unit (CFU) counting, reduction in optical density (OD) in co-cultures and assay of killing efficiency were carried out. Also, the raw wastewater was treated by Bdellovibrio then the reduction in CFU counting and reduction in OD was evaluated.Results:Four strains of Bdellovibrio were isolated and were registered in Gene Bank. Clear plaques were observed after 3–6 days of incubation for all prey cells. The CFU enumerations of all preys were decreased after 48 hrs in co-cultures and raw wastewater. Also, OD was decreased down to 0.2 nm after 48 hrs.Conclusion:These autochthonous Bdellovibrio strains are proposed to use for bio-control of Gram-negative pathogenic bacteria in wastewater and reuse it for irrigation in arid regions.

Complete Genome Sequence of Bdellovibrio sp. Strain KM01, a Predatory Bacterium Isolated from Soil

We report the complete genome sequence of the predatory bacterium Bdellovibrio sp. strain KM01, isolated from soil collected near a pond. The genome is 3,961,288 bp long with 45.5% GC content. Comparative genomics among Bdellovibrio strains will help us understand how genotypic differences affect differences in predatory phenotypes.

Isolation of Bdellovibrio sp. from Wastewater and Their Potential Application in Control of Salmonella paratyphi in Water

ABSTRACTThe problem of the increasing resistance of bacteria to conventional antibiotics gives the bacteria Bdellovibrio a great utility as a potential alternative source of antibiotics. Therefore, the preliminary goal of the present study was isolation and identification of antibiotic-resistant bacteria used as prey organisms for isolated Bdellovibrio sp., by xylose lysine desoxycholate (XLD) agar from different types of water in the Taif area, Saudi Arabia, and also to investigate water quality. Four antibiotic-resistant isolates of Salmonella sp. which were susceptible to Bdellovibrio were identified by morphological, biochemical and 16S rRNA characterization as Salmonella paratyphi. Seventeen strains of Bdellovibrio sp. were isolated from sewage wastewater using isolated S. paratyphi as prey bacteria by a double-layer plate. Only one of them causing a large plaque after 48 h of incubation at 37°C was designated Bdellovibrio AOA12. The shape of Bdellovibrio was confirmed by morphological characterization and electron microscopy. Bdellovibrio could lyse four antibiotic-resistant Gram-negative bacterial strains of Salmonella paratyphi but could not lyse Gram-positive bacteria such as Staphylococcus aureus and Bacillus cereus. The kinetic lysis of the Bdellovibrio as predator to four isolates of antibiotic-resistant Salmonella paratyphi as prey organisms was demonstrated. The results suggest that it may be possible to utilize Bdellovibrio to control antibiotic-resistant S. paratyphi in water.

Isolation of Bdellovibrio sp. from soil samples in Mexico and their potential applications in control of pathogens.

In this study, two strains of Bdellovibrio were isolated from soil samples using the culture‐dependent technique and two members of the family Enterobacteriaceae (Klebsiella sp. and Salmonella sp.) as prey. The Bdellovibrio strains were bacteriolytic, plaque‐forming, and highly motile gram‐negative bacteria. We identified and confirmed the Bdellovibrio strains using microscopy, PCR amplification, and sequencing of the 16S rRNA gene. They were observed to be different strains based on hit locus and prey range analyses. Here, the first report on Bdellovibrio strains isolated from soil in Mexico corroborates earlier report indicating that populations of Bdellovibrio found in soil are heterogeneous thereby the need to identify the various strains.

Genome-wide comparative analysis of ABC systems in the Bdellovibrio-and-like organisms

Bdellovibrio-and-like organisms (BALOs) are gram-negative, predatory bacteria with wide variations in genome sizes and GC content and ecological habitats. The ATP-binding cassette (ABC) systems have been identified in several prokaryotes, fungi and plants and have a role in transport of materials in and out of cells and in cellular processes. However, knowledge of the ABC systems of BALOs remains obscure. A total of 269 putative ABC proteins were identified in BALOs. The genes encoding these ABC systems occupy nearly 1.3% of the gene content in freshwater Bdellovibrio strains and about 0.7% in their saltwater counterparts. The proteins found belong to 25 ABC system families based on their structural characteristics and functions. Among these, 16 families function as importers, 6 as exporters and 3 are involved in various cellular processes. Eight of these 25 ABC system families were deduced to be the core set of ABC systems conserved in all BALOs. All Bacteriovorax strains have 28 or less ABC systems. On the contrary, the freshwater Bdellovibrio strains have more ABC systems, typically around 51. In the genome of Bdellovibrio exovorus JSS (CP003537.1), 53 putative ABC systems were detected, representing the highest number among all the BALO genomes examined in this study. Unexpected high numbers of ABC systems involved in cellular processes were found in all BALOs. Phylogenetic analysis suggests that the majority of ABC proteins can be assigned into many separate families with high bootstrap supports (>50%). In this study, a general framework of sequence–structure–function connections for the ABC systems in BALOs was revealed providing novel insights for future investigations.

Genome analysis of a simultaneously predatory and prey-independent, novel Bdellovibrio bacteriovorus from the River Tiber, supports in silico predictions of both ancient and recent lateral gene transfer from diverse bacteria

BackgroundEvolution equipped Bdellovibrio bacteriovorus predatory bacteria to invade other bacteria, digesting and replicating, sealed within them thus preventing nutrient-sharing with organisms in the surrounding environment. Bdellovibrio were previously described as “obligate predators” because only by mutations, often in gene bd0108, are 1 in ~1x107 of predatory lab strains of Bdellovibrio converted to prey-independent growth. A previous genomic analysis of B. bacteriovorus strain HD100 suggested that predatory consumption of prey DNA by lytic enzymes made Bdellovibrio less likely than other bacteria to acquire DNA by lateral gene transfer (LGT). However the Doolittle and Pan groups predicted, in silico, both ancient and recent lateral gene transfer into the B. bacteriovorus HD100 genome.ResultsTo test these predictions, we isolated a predatory bacterium from the River Tiber- a good potential source of LGT as it is rich in diverse bacteria and organic pollutants- by enrichment culturing with E. coli prey cells. The isolate was identified as B. bacteriovorus and named as strain Tiberius. Unusually, this Tiberius strain showed simultaneous prey-independent growth on organic nutrients and predatory growth on live prey. Despite the prey-independent growth, the homolog of bd0108 did not have typical prey-independent-type mutations. The dual growth mode may reflect the high carbon content of the river, and gives B. bacteriovorus Tiberius extended non-predatory contact with the other bacteria present. The HD100 and Tiberius genomes were extensively syntenic despite their different cultured-terrestrial/freshly-isolated aquatic histories; but there were significant differences in gene content indicative of genomic flux and LGT. Gene content comparisons support previously published in silico predictions for LGT in strain HD100 with substantial conservation of genes predicted to have ancient LGT origins but little conservation of AT-rich genes predicted to be recently acquired.ConclusionsThe natural niche and dual predatory, and prey-independent growth of the B. bacteriovorus Tiberius strain afforded it extensive non-predatory contact with other marine and freshwater bacteria from which LGT is evident in its genome. Thus despite their arsenal of DNA-lytic enzymes; Bdellovibrio are not always predatory in natural niches and their genomes are shaped by acquiring whole genes from other bacteria.

Characterization of outer membrane protein fractions of Bdellovibrionales

Bdellovibrio-and-like organisms (BALOs) are predatory bacteria that prey upon Gram-negative bacteria and are taxonomically subsumed in the order Bdellovibrionales. Despite their unique lifestyle, these bacteria show remarkable genotypic diversities. The outer membrane of the predators is likely to play an important role during the recognition and invasion stage, as well as in the intraperiplasmic growth phase. In this study, the outer membrane protein fractions of type strains of Bdellovibrio, Bacteriovorax and Peredibacter were investigated, revealing the presence of outer membrane proteins (Omps) similar to the major Omps of Bdellovibrio bacteriovorus. The primary structures of these Omps of Bdellovibrio sp. W, Bacteriovorax stolpii and Peredibacter starrii were elucidated by a combined mass spectrometric–reverse genetic approach. The similarity between the analyzed Omps of the investigated BALOs ranges from 32% to 89% showing conserved amino acid regions in their primary structure.

Diversity and evolution of Bdellovibrio-and-like organisms (BALOs), reclassification of Bacteriovorax starrii as Peredibacter starrii gen. nov., comb. nov., and description of the Bacteriovorax-Peredibacter clade as Bacteriovoracaceae fam. nov.

A phylogenetic analysis of Bdellovibrio-and-like organisms (BALOs) was performed. It was based on the characterization of 71 strains and on all consequent 16S rRNA gene sequences available in databases, including clones identified by data-mining, totalling 120 strains from very varied biotopes. Amplified rDNA restriction analysis (ARDRA) accurately reflected the diversity and phylogenetic affiliation of BALOs, thereby providing an efficient screening tool. Extensive phylogenetic analysis of the 16S rRNA gene sequences revealed great diversity within the Bdellovibrio (> 14 % divergence) and Bacteriovorax (> 16 %) clades, which comprised nine and eight clusters, respectively, exhibiting more than 3 % intra-cluster divergence. The clades diverged by more than 20 %. The analysis of conserved 16S rRNA secondary structures showed that Bdellovibrio contained motifs atypical of the delta-Proteobacteria, suggesting that it is ancestral to Bacteriovorax. While none of the Bdellovibrio strains were of marine origin, Bacteriovorax included separate soil/freshwater and marine-specific groups. On the basis of their extensive diversity and the large distance separating the groups, it is proposed that Bacteriovorax starrii be placed into a new genus, Peredibacter gen. nov., with Peredibacter starrii A3.12T (= ATCC 15145T = NCCB 72004T) as its type strain. Also proposed is a redefinition of the Bdellovibrio and the Bacteriovorax-Peredibacter lineages as two different families, i.e. 'Bdellovibrionaceae' and a new family, Bacteriovoracaceae. Also, a re-evaluation of oligonucleotides targeting BALOs is presented, and the implications of the large diversity of these organisms and of their distribution in very different environments are discussed.

Taxonomic Studies of Predatory Bdellovibrios Based on 16S rRNA Analysis, Ribotyping and the hit Locus and Characterization of Isolates from the Gut of Animals

The aim of our study was to obtain data for the molecular characterization of bdellovibrio bacteria, which were recently split into the genus Bdellovibrio and the newly designated genus Bacteriovorax. We determined the 16S rDNA sequences of five reference strains and performed a phylogenetic analysis including published 16S rRNA sequences of bdellovibrios. A comparison of the secondary structure showed significant differences in two regions of the 16S rRNAs of the species Bdellovibrio bacteriovorus, Bacteriovorax starrii, and Bacteriovorax stolpii. In addition, ribotyping techniques gave specific hybridization patterns and revealed that two rRNA operons are present in the investigated strains. A hybridization probe derived from the genetic locus hit, associated with the host independent (HI) phenotype of B. bacteriovorus, was found to be specific for this species. Sequence comparison of the hit locus revealed few base pair changes between host independent (HI) and host dependent (HD) strains. Ribotyping and hybridization experiments using the hit probe were applied to characterize bdellovibrio strains isolated from the gut of animals and humans and one isolate from sewage.

A comparison of the survival of intraperiplasmic and attack phase bdellovibrios with reduced oxygen

The ability of intraperiplasmic and attack phase bdellovibrios to survive and/or grow under anoxic and microaerobic conditions was examined. Both halotolerant and nonhalotolerant bdellovibrio strains were examined. In all instances, the bdellovibrio strains were unable to grow under anoxic conditions, but were able to survive for periods of time in both the extracellular and intraperiplasmic forms. However, the intraperiplasmic organisms were observed to survive longer. Increased temperature hastened the loss of viability of both forms of the predatory bacteria in oxic and anoxic environments. Under microaerobic conditions, halotolerant bdellovibrios were observed to grow, although at a slightly reduced rate than in atmospheric oxygen, while two nonhalotolerant isolates survived but did not grow. The ability of attack phase bdellovibrios to survive in an anoxic environment for up to nine days and their growth or survival under microaerobic conditions greatly expands the possible ecological niches in which the predators may be active members of the microbial community.

Ecology and characteristics of bdellovibrios from three tropical marine habitats

Marine bdellovibrios have not previously been reported from the southern hemisphere, and knowledge of their occurrence in marine ecosystems is rudimentary. This study examined quantitative and qualitative aspects of bdellovibrios parasitic to the bacterium Vibrio alginolyticus at each of three representative tropical marine habitats of the Great Barrier Reef. Bdellovibrios were found in the water column throughout a 12 mo period from May 1992 at a sandy beach, a mangrove and a fringing coral reef. Their abundance was correlated with water temperature (P 0.05). They were more abundant in sub-surface than bottom waters in summer, but the reverse occurred in winter. Midwater samples usually had least bdellovibrios. Bdellovibrio numbers were significantly correlated with those of potential host bacteria—colony-forming bacteria at all habitats and total bacteria at the beach and reef habitats. Strain characteristics, primarily based on host range, indicated qualitative differences in bdellovibrio populations among habitats. Pseudomonas atlantica, P. aeruginosa, P. marina, Cytophaga marinoflava, Vibrio gazogenes, V. mimicus and a Spirillum-like bacterium were not parasitised by bdellovibrios from any habitat. Of the other 25 Vibrio spp. tested, most were parasitised by the majority of bdellovibrio strains from each habitat. Strain differences were principally with respect to parasitism of non-Vibrio bacteria. All strains required Na+ and grew at 35°C, but some failed to grow at 15°C.

Relationships among the bdellovibrios revealed by partial sequences of 16S ribosomal RNA

Members of the genusBdellovibrio possess the unifying phenotypic trait of attacking and preying upon other Gram-negative bacteria. It has been suggested that this common lifestyle arose by convergent evolution. Physiological and G + C studies have led to the notion that bdellovibrios are a heterogeneous group of loosely related bacteria. We have inferred the phylogenetic relatedness of 12 strains ofBdellovibrio through the analysis of partial 16S ribosomal RNA sequences. Similarity and degree of homology were assessed, and a phylogenetic tree was constructed by the distance matrix method. One branch of the two-branched tree consisted ofB. bacteriovorus and related strains (W, 6-5-S, 109, 109D, 109J, 114, HI Ox9-2, and HI Ox9-3). The other branch was itself branched, withB. starrii, B. stolpii, and marine strain BM4 in separate sub-branches. AllBdellovibrio strains in turn clustered with representatives of the delta division of theProteobacteria. The results indicate that there are at least two subdivisions of the genusBdellovibrio and that present-day bdellovibrios arose from a common ancestor. The placement of the genusBdellovibrio within the delta division of theProteobacteria was confirmed.

Isolation and characterization of marine and salt pond halophylic bdellovibrios

A method to enrich and isolate bdellovibrios from marine waters and solar salt evaporation ponds of a salinity up to 15.5% has been developed. The method promotes growth of gram-negative autochthonous bacteria followed by bdellovibrios that prey on them. Various Vibrio and Pseudomonas strains have been used to reveal Bdellovibrio plaques and in no case have bacteriophages interfered with the isolation technique. A total of 13 strains have been characterized by electron microscopy, host range, cytochrome spectra, and capacity to grow at different salt concentrations. All isolates show typical Bdellovibrio morphology and life cycle. Little differences could be found among the Bdellovibrio strains according to host range patterns with the exception of Bdellovibrio BS42 and BS43 that were unable to lyse various of the hosts used. These two Bdellovibrio strains differed too from the others in the cytochrome spectra of whole cell suspensions (α and β bands of the cytochrome c). All of the bdellovibrios isolated showed the typical bands of cytochromes a and c, and in some cases, the peak of cytochrome b could also be discerned. The Soret band has proved to be of great value to distinguish halophylic from freshwater bdellovibrios. The saline growth range found in this study is broader than the one described previously for marine bdellovibrios (1–6%). Some of the strains isolated from seawaters or from salt ponds could grow from 1 to 15% total salts salinity. The growth capacity at the highest salinities depends to a great extent on the proportion of salts in the medium. A compensated saline solution supports better Bdellovibrio proliferation than a solution where only the NaCl concentration has been increased. Tolerance to salinity divides the isolated bdellovibrios into two groups: a low salinity group of strains of marine origin able to grow at a salinity of 1% total salts, but not in freshwater media nor above 12% total salts, and a high salinity group isolated from solar evaporation ponds, with strains able to multiply in liquid media up to a salinity of 15% total salts, but unable to grow at a salinity of 1%. These results broaden the habitat of Bdellovibrio in saline waters of marine origin up to salinities of 15%, as well as indicate further diversity in this group of bacterial predators.Key words: halophylic Bdellovibrio, salterns, marine bacteria.

Verification of the protein in the outer membrane of Bdellovibrio bacteriovorus as the OmpF protein of its Escherichia coli prey.

Two research groups showed that several Bdellovibrio strains incorporated into their outer membranes intact OmpF porin proteins derived from their Escherichia coli prey. These results could not be reproduced by another group using Bdellovibrio bacteriovorus 109J. They showed that a major protein appearing in the Bdellovibrio Triton X-100-insoluble outer membrane was coded for by the bdellovibrios. We reconciled these results by examining the strain used by this group and by reviving a freeze-dried culture of strain 109J which had been stored for almost 9 years. B. bacteriovorus 109J failed to acquire substantial amounts of the OmpF protein from E. coli ML35, and a protein coded for by the bdellovibrios was expressed in its place. However, B. bacteriovorus 109J incorporated the OmpF protein from rough K-12 strains of E. coli, and the revived 9-year-old culture of B. bacteriovorus 109J incorporated more of the OmpF protein from the smooth E. coli ML35 than did its contemporary counterpart. The protein isolated from the outer membrane of the bdellovibrios was identified as the OmpF protein of E. coli by its protease peptide profile on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and by Western blot analysis. This confirmed that bdellovibrios relocalize outer membrane proteins from their prey, but relocalization may be an unstable trait which can be influenced by the prey.

Serotyping of Bdellovibrios by Agglutination and Indirect Immunofluorescence

A comparative serological study was undertaken to clarify the uncertain interrelationships among various bdellovibrios. A total of 17 predacious (hostdependent) strains and 12 nonpredacious strains were serotyped. Strains were grouped into serotypes on the basis of positive agglutination and strongly positive fluorescence cross-reactions. The results of division of the strains into serogroups by these two methods were identical. Bdellovibrio starrii strain A3.12 and Bdellovibrio stolpii strain UKi2 were found to be antigenically distinct from each other and from the group of strains comprising Bdellovibrio bacteriovorus; the latter was found to be an antigenically heterogeneous group consisting of at least nine serogroups. All nonpredacious strains were found to be related antigenically to their obligately predacious counterparts. An antigen(s) common to all of the Bdellovibrio strains examined was exhibited as weakly positive fluorescence. Selected strains of Bdellovibrio were studied by Ouchterlony immunodiffusion, which confirmed the serogrouping results and provided a technically simple method of serotyping. Further immunochemical characterization of the strains of Bdellovibrio in conjunction with increased knowledge of other differences among strains will likely result in the formation of additional species of Bdellovibrio.

腸炎ビブリオ寄生溶菌性Ｂｄｅｌｌｏｖｉｂｒｉｏの生理生態に関する研究

A host sea water(HS) agar plate method for the isolation and enumeration of marine Bdellovibrio, parasitic and lytic to Vibrio parahaemolyticus, was developed in this work. A few Bdellovibrio strains were isolated from coastal sea water and the physiological multiplication phase, similar to that of common bacteria, was observed. The raw sea water samples were inoculated with about107/ml cells of V. parahaemolyticus (Biotype 1) and the related organisms (Biotype 2 and 3), respectively, and incubated at 25°C. After 2-4 days of incubation, population of these bacteria decreased approximately to 104/ml, whereas temporal multiplication of the Bdellovibrio was observed. Using HS agar plate prepared with the Biotype 1, 2 or 3, respectively, 26 Bdellovibrio strains were isolated from coastal waters. These isolates showed highest lytic activity to Vibrio and Pseudomonas organisms among the marine bacterial flora. Lytic action of the isolates to the various terrestrial bacteria varied with the Bdellovibrio strains. The occurrence of some Bdellovibrio resistant strains was observed among the Kanagawa phenomenon negative V. perahaemolyticus.

Chemotaxis of Bdellovibrio bacteriovorus toward pure compounds.

Positive chemotaxis by Bdellovibrio bacteriovorus strain UKi2 was measured for 139 compounds. Twenty-one compounds were attractants; sensitive attraction was elicited by acetate, propionate, thioacetate, malonate, cis-oxalacetate, D-glucose-6-phosphate, acetyl coenzyme A, ammonium ion, barium ion, manganous ion, and potassium ion. Several of the attractants for B. bacteriovorus strain UKi2 also were attractants to strains 6-5-S and 114; however, strains 109D and 109J were not attracted by the compounds tested. Of 33 compounds tested, 8 were repellents for B. bacteriovorus strain UKi2: n-caproate, alanine, isoleucine, leucine, phenylalanine, tyrosine, cobaltous chloride, and hydronium ion. None of the organic repellents for strain UKi2 elicited repulson from strains 114 or 109D. However, all three strains of Bdellovibrio show aerotaxis. Several compounds were tested for their effects on viability and predacious growth of B. bacteriovorus strain UKi2. No simple correlation was found between attraction or repulsion and benefit or harm to bdellovibrios. The data are consistent with the view that in nature, the greatest survival value of chemotaxis for bdellovibros may be in aerotaxis, attraction to certain inorganic ions and acetate, and repulsion by hydronium ion.

Attempts to grow bdellovibrios micurgically-injected into animal cells

Incubation in buffer of Bdellovibrio bacteriovorus 109J, B. stolpii UKi2, or B. starrii A3.12 with washed eucaryotic animal cells (mouse liver, hamster kidney, or bovine mammary gland) resulted in neither attachment nor growth of the bdellovibrios. When cells of these bdellovibrio strains were incubated with erythrocyte suspensions (bovine or rabbit) a very low level of bdellovibrio attachment and penetration occurred, but no growth could be detected. Using micurgical procedures, bdellovibrios were injected into the perivetelline space or the cytoplasm of rabbit ova. After 18–24h incubation, neither a significant loss nor increase of injected, intracellular bdellovibrios was observed. Limited axenic growth of bdellovibrios (109J or UKi2) occurred in media containing rabbit ova extracts and dilute nutrient broth. It is concluded that eucaryotic rabbit ova do not provide a suitable environment for intracellular bdellovibrio growth.

Isolation, enumeration, and host range of marine Bdellovibrios.

Thirteen strains of Bdellovibrio were isolated from sea water off the coast of Oahu, Hawaii. Electron microscopic examination of the isolates indicated that all had a single, sheathed, polar flagellum with a diameter of 24–27 nm. Determination of host range on species of marine bacteria (Beneckea, Photobacterium, Alteromonas, Pseudomonas, and Alcaligenes) as well as terrestrial species (Escherichia, Salmonella, Aerobacter, Aeromonas, Vibrio, and Pseudomonas) indicated that the marine bdellovibrios were able to parasitize 17–32 species out of a total of 42 tested. In general, bacteria of marine origin were better hosts than terrestrial strains; Vibrio cholerae, Aeromonas formicans, and Salmonella typhimurium were, however, lysed by some strains of Bdellovibrio at a rate and intensity comparable to the marine hosts. A numerical analysis of the host range indicated that on the basis of this property the 13 strains of Bdellovibrio could be subdivided into three groups. Three HI strains were isolated from one group and were found to have a moles % GC content in their DNA of 38.6 moles %, different from that of any described terrestrial species. The moles % GC content of a HI strain from another group was 43.5. All of the marine isolates required 75–100 mM sodium chloride for plaque formation and the optimum was 125–150 mM. The requirement for sodium could not be replaced by potassium. HI strains grew only in media in which sodium was present and failed to grow in media in which this ion was replaced by equimolar amounts of potassium. Enumeration studies indicated that the coastal waters of Oahu, Hawaii, had about 121–194 bdellovibrios per liter of sea water. The results of this study suggested that the marine bdellovibrios differed from terrestrial isolates in a number of properties which were a reflection of their marine habitat.

Isolation and preliminary characterization of bacteriophages for Bdellovibrio bacteriovorus.

Ten bacteriophages that attack and lyse saprophytic strains of Bdellovibrio bacteriovorus were isolated. Morphological, serological, and host-range studies revealed that there were four different bdellovibrio phages present among the isolates. One of the phages lysed a strain of B. bacteriovorus that requires the presence of a suitable bacterial host for growth. The phage attached to the bdellovibrio cells in the absence of the bacterial host cells; lysis occurred only in the presence of host cells. The 19 saprophytic bdellovibrio strains employed in the phage host-range studies were grouped on the basis of their susceptibility to phage lysis.