Host specificity of Salmonella weltevreden typing phages

The host range of the six S. weltevreden typing phages was studied on 1469 strains belonging to 37 different Salmonella serotypes. In addition to S. weltevreden, only S. nchanga, S. give, S. lexington and S. anatum, all belonging to O group E₁, showed varying degrees of susceptibility to the action of some of the typing phages.Typing phage VI lysed only one strain other than S. weltevreden. All serotypes tested other than S. weltevreden were resistant to phages III and IV even at 1000 times the routine test dilution. Thus, typing phages III and IV were specific for S. weltevreden. The sensitivity patterns of S. weltevreden typing phages were not found to bear much correlation with either somatic of flagellar antigens of Salmonellae.     

Phage specificity and lipopolysaccarides of stem- and root-nodulating bacteria (Azorhizobium caulinodans, Sinorhizobium spp., and Rhizobium spp.) of Sesbania spp

Phage susceptibility pattern and its correlation with lipopolysaccharide (LPS) and plasmid profiles may help in understanding the phenotypic and genotypic diversity among highly promiscuous group of rhizobia nodulating Sesbania spp.; 43 phages were from two stem-nodulating bacteria of S. rostrata and 16 phages were from root-nodulating bacteria of S. sesban, S. aegyptica and S. rostrata. Phage susceptibility pattern of 38 Sesbania nodulating bacteria was correlated with their LPS rather than plasmid profiles. Different species of bacteria (A. caulinodans- ORS571, SRS1-3 and Sinorhizobium saheli- SRR907, SRR912) showing distinct LPS subtypes were susceptible to different group of phages. Phages could also discriminate the strains of Si. saheli (SSR312, SAR610) possessing distinct LPS subtypes. Phages of Si. meliloti (SSR302) were strain-specific. All the strains of R. huautlense having incomplete LPS (insignificant O-chain) were phage-resistant. In in vitro assay, 100% of the phages were adsorbed to LPS of indicator bacterium or its closely related strain(s) only. These observations suggest the significance of LPS in phage specificity of Sesbania nodulating rhizobia. Highly specific phages may serve as biological marker for monitoring the susceptible bacterial strains in culture collections and environment.     

Bacteriophage typing of Salmonella weltevreden

Salmonella weltevreden has been found to be one of the commonest Salmonella serotypes isolated from diverse sources in India and has also been isolated in a number of other countries. A phage typing scheme was developed for this serotype using a set of six typing phages. These phages had been selected out of 146 phage strains isolated and purified from stool samples of man, laboratory animals and other animals, sewage and surface water sources, and the lytic mutants of temperate phages from S. weltevreden.The phage typing scheme was applied systematically to type the 946 strains from India isolated during 1958–1974 and 148 strains originating from Australia, Burma, England, Gan Island, Holland, Hong Kong, Malaysia, New Zealand, Papua New Guinea, The Philippines, Thailand, The United States and Vietnam during 1953–1971. The scheme was particularly studied to evaluate its utility in mapping the epidemiologically related strains from various sources.The S. weltevreden strains could be classified into ten phage types. Phage types 2 and 7 were found exclusively amongst Indian strains, type 6 from Vietnam and type 8 from Burma, Thailand and Vietnam. Phage types were found to be stable and consistent with the independent epidemiological data available.     

Lysogenic strains of lactic Acid streptococci and lytic spectra of their temperate bacteriophages

A total of 113 strains of mesophilic strains lactic streptococci of the species Streptococcus lactis, S. lactis subsp. diacetilactis, and S. cremoris, chosen from 291 strains that had been previously classified into six groups on the basis of their sensitivity to 132 virulent phages, were subjected to induction with mitomycin C. Among these strains, 43% produced phages capable of forming plaques of lysis on an indicator strain either spontaneously or after induction. There was a close correlation between the lytic spectra of temperate and virulent phages. Among the strains studied, 25% were shown to be indicator strains. These results emphasized the high probability of development of temperate phages in a starter culture containing mesophilic lactic streptococci and therefore their importance as a cause of accidents in cheese making.     

Studies of natural populations and mutants of rhizobium in the improvement of legume inoculants

Summary A study has been made of the symbiotic effectiveness ofRhizobium trifolii in fields ofTrifolium subterraneum in south-eastern Australia, with the purpose of providing background information for a programme of inoculant strain improvement. The strains found varied widely in symbiotic effectiveness. The distribution patterns of effectiveness varied from year to year and from locality to locality. From 24 to 65% of strains ofR. trifolii from different localities exhibited antagonism to selectedR. trifolii indicator strains. These reactions were mainly due to mild antibiotic effects but bacteriocinogenic strains and strains producing specific, virulent or wide-range, temperate phages, were also common. Factors to be considered in the selection and evaluation of inoculant strains from among natural populations are discussed and the possible role of genetic manipulation is examined. It is concluded that the general application of DNA transformation and transduction is restricted because of the limited degree of DNA homology among strains, the limited host-range of transducing phages and the lack of suitablein vitro screening procedures for symbiotic characters. A mutational model is presented in which characters known to be associated with symbiotic effectiveness would be manipulated by mutation and back-mutation to effect quantitative increases in effectiveness.     

Lysogeny among mycobacteria

Our investigations to detect naturally lysogenic strains of mycobacteria were limited to 1 strain ofMycobacterium smegmatis, 4 strains ofMycobacterium borstelense var.niacinogenes, and to 5 strains ofMycobacterium marinum (Syn:Mycobacterium balnei), all together 10 strains. They were chosen because as a sign of lysis they secrete a large quantity of cytoplasmatic components (nucleic acids proteins, amino acids etc.) into the fluid medium (for instance phosphate buffer), in which they are suspended. In a first series of experiments culture filtrates were tested on 84 strains of slowly and rapidly growingMycobacterium species as indicator strains. Using this method free phage particles were only found in the culture filtrate of 1 strain,Mycobacterium smegmatis SN 46, isolated from a patient with achalasia. Phage particles could not be found in the filtrates of the other 9 probably lysogenic strains. In a second series of experiments more closely related indicator strains were used. The 10 probably lysogenic strains were cultured in bovine serum or antiphage-antiserum containing medium and single selected colony cultures a small part of which showed sensitivity to the filtrates. The released and adapted phages, designated as B₂₄, B₃₀, B₃₂, B₃₃, B₃₄ and B₃₅ have a very narrow host range. The plaques are very small and turbid. On electron micrographs the temperate phages B₂₄, B₃₀ and B₃₅ exhibit the typical head-tail morphology. The head of the temperateborstelense var.niacinogenes phage B₃₀ is 45 nm in diameter, the tength of tail is about, 120nm. The average dimensions of the long head ofsmegmatis phage B₂₄ are 40 × 80 nm, the tail is about 160 nm long. The balnei phage B₃₅ is very similar morphologically to phage B₃₀. The head is about 50 nm in diameter, the length of tail about 160 nm. The phage sensitive variants are not “carrier” strains. Their phage sensitivity is not a stable property. After several culture passages in serum-free medium the variants regain their phage immunity completely and release phages like the lysogenic parent strains. The sensitive variants must therefore be considered to be also lysogenic. TheMycobacterium borstelense var.niacinogenes phages are serologically very related. Dedicated to Academician Ivan Málek on the occasion of his 60th birthday     

Biological characteristics ofSalmonella weltevreden typing phages

The important biological characteristics ofSalmonella weltevreden (3,10∶r∶z₆) typing phages were studied. On the basis of these, the phages could be classified into three groups: phages Φ I and Φ II, phages Φ III, Φ IV and Φ VI, and phage Φ V. Part of the work was done at the National Salmonella and Escherichia Centre, Central Research Institute, Kasauli, India.     

Phage Typing Scheme for Salmonella braenderup

A phage typing scheme for Salmonella braenderup based on both phage sensitivity and lysogenicity patterns is presented. Three S. braenderup symbiotic phages (br A, br B, and br C) were used in the phage sensitivity test, and three indicator strains were used for the lysogenicity test. The 424 strains examined were grouped in 15 of 64 possible types within this scheme. The most frequent types were: A(1) (32%), G(5) (21%), F(2) (14%), and H(1) (8%). All of the strains isolated successively from the same person belonged to the same type, as did the strains isolated from groups of individuals related to a common source.     

Lysogenicity of atypical strains ofSalmonella paratyphi B

Summary Atypical strains ofS. paratyphi B have been described which caused gastroenteritis or were found by chance in healthy perons. These strains possessed atypical natural phages. The late fermentation of d-tartrate by some strains ofS. paratyphi B was examined.     

Phage-typing ofSalmonella weltevreden based on lysogeny II. Epidemiological usefulness of the system and geographical distribution of its phage-types

Nine hundred and forty-six strains ofSalmonella weltevreden isolated in different states of India during 1958–1974 and 124 strains from Australia, Burma, Holland, Hong Kong, New Zealand, Papua New Guinea, the Philippines, Thailand, the United States and Vietnam during 1953–1971 were phagetyped according to the phage-typing scheme described in the first part of this paper (Sood and Basu, 1977). The epidemiological incidence and geographical distribution of phage-types ofSalmonella weltevreden were studied. All the phage-types were present in India, the predominant phage-types being b, d and i. Phage-type g was isolated exclusively from India. All the 14 strains from Hawaii belonged to phage-type i. Phage-type h was the most predominant phage-type in Vietnam. The 15 strains isolated from Papua New Guinea in 1965, which were supposed to have originated from a single source, belonged to 3 phage-types. Except these cultures all the available epidemiologically related strains were of uniform phage-types — a finding which establishes the epidemiological validity of the scheme.     

Phage susceptibility and plasmid profile analysis of Sinorhizobium fredii

This is the first report identifying bacteriophages and documenting megaplasmids of Sinorhizobium fredii. Plasmid DNA content and bacteriophage typing of eighteen strains of S. fredii were determined. S. fredii strains fell into ten plasmid profile groups containing 1 to 6 plasmids, some evidently larger than 1000 MDa. Twenty-three S. fredii lytic phages were isolated from soil, and they lysed six different S. fredii strains. The host range and plaque morphology of these phages were studied. Susceptibility to S. fredii phages was examined for S. meliloti; Rhizobium leguminosarum bvs. viceae, trifolii and Phaseoli; R. loti; Bradyrhizobium japonicum; B. elkanii and Bradyrhizobium sp. (Arachis). Several phages that originally lysed S. fredii strain USDA 206 also lysed strains of all three S. fredii serogroups described originally by Sadowsky et al. Phages that infected S. fredii strains USDA 191 and USDA 257 were highly specific and lysed only serogroup 193 strains. S. meliloti strains L5-30 and USDA 1005 were lysed by three of the phages that lysed S. fredii strain USDA 217. No other Rhizobium or Bradyrhizobium strain tested was susceptible to lysis by any of the S. fredii phages. The present investigation indicates that phage susceptibility in conjunction with plasmid profile analysis may provide a rapid method for identification and characterization of strains of S. fredii.     

Investigation of the lytic ability of South African bacteriophages specific for Staphylococcus aureus,associated with bovine mastitis

Bovine mastitis is an infectious disease of the mammary glands of dairy cattle primarily causaled by the bacterium, Staphylococcus aureus subsp. aureus Rosenbach1884. Traditional control of this organism was through the use of antibiotics. However, S. aureus is developing resistance towards these chemotherapeutic agents faster than they are being developed. Bacteriophages can serve as an alternative control measure for the disease. This study investigated the prevalence of phages and S. aureus within the South African dairy environment, as well as infectivity of phage isolates against antibiotic-resistant S. aureus. The four S. aureus strains used in the study displayed resistance to representative antibiotics from both the β-lactamases and non-β-lactamases, macrolides, aminoglycosides and glycopeptides. Susceptibility was only noted towards the tetracycline antibiotics. Twenty-eight phages were isolated and screened against four strains of S. aureus. Only six phages showed biocontrol potential based on their wide host range, high titres and common growth requirements. Morphological and preliminary genomic analysis was carried out on the three best performing phages. At an optimal titre of between 6.2 × 10⁷ and 2.9 × 10⁸ pfu.ml^?1, the phages were able to reduce live bacterial cell counts between 64% and 95%. In addition, these six phages showed further infectivity towards S. aureus strains that were isolated from different milk-producing regions during a farm survey. The phages isolated in this study show reasonable potential for in vivo applications.     

A sub-division ofSalmonella typhimurium into phage types based on the method of craigie and yen; Phages adaptable to species of the B and D group ofSalmonella; Phase adsorption as diagnostic aid

Summary and conclusions Phages were isolated which were adsorbed bySalmonella strains of groups B and D. These phages were adaptable to strains of species from these groups. The behaviour of some adaptations of these phages on strains ofS. paratyphi B has been described. A typing scheme forS. typhimurium has been established. So far, this consists of 32 types which have all been demonstrated with adaptations of a single phage.     

Lysogeny in Lactobacillus delbrueckii strains and characterization of two new temperate prolate-headed bacteriophages

Aims: Frequency of lysogeny in Lactobacillus delbrueckii strains (from commercial and natural starters) and preliminary characterization of temperate bacteriophages isolated from them. Methods and Results: Induction of strains (a total of 16) was made using mitomycin C (MC) (0·5 μg ml⁻¹). For 37% of the MC-treated supernatants, it was possible to detect phage particles or presence of killing activity, but only two active bacteriophages were isolated. The two temperate phages isolated were prolate-headed phages which belonged to group c of Lact. delbrueckii bacteriophages classification. Different DNA restriction patterns were obtained for each phage, while the structural protein profiles and packaging sites were identical. Distinctive one-step growth curves were exhibited by each phage. An influence of calcium ions was observed for their lysis in broth but not on the adsorption levels. Conclusions: Our study showed that lysogeny is also present in Lact. delbrueckii strains, including commercial strains. Significance and Impact of the Study: Commercial strains could be lysogenic and this fact has a great practical importance since they could contribute to the dissemination of active-phage particles in industrial environments.     

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.     

Lysogeny in encapsulated and nontypable strains ofHaemophilus influenzae

Encapsulated and nontypableHaemophilus influenzae isolates recovered from pediatric patients and healthy children were examined for their ability to lyse and to release phage after mitomycin C induction. Lysis occurred in 16 out of 58 isolates tested for lysogeny. The serotype b capsule ofH. influenzae does not exhibit an inhibitory effect on either cell lysis or the ability of the cells to become lysogenized. Electron microscopic examination revealed the presence of tailed particles in 11 lysates. The tailed particles belong to two morphological groups (A and B) according to the classification of Bradley [Bacteriol. Rev. 31:230–314]. None of the phages was able to produce plaques when tested on a large number of strains. However, five lysates exhibited killing properties involving several nontypableH. influenzae strains. This effect sedimented with the bacteriophage after centrifugation. All the phages recovered fromH. influenzae isolates in this study appeared to be genetically defective.     

Toward rational control of<Emphasis Type="Italic"> Escherichia coli</Emphasis> O157:H7 by a phage cocktail

Twenty six phages infected with Escherichia coli O157:H7 were screened from various sources. Among them, nine caused visible lysis of E. coli O157:H7 cells in LB liquid medium. However, prolonged incubation of E. coli cells and phage allowed the emergence of phage-resistant cells. The susceptibility of the phage-resistant cells to the nine phages was diverse. A rational procedure for selecting an effective cocktail of phage for controlling bacteria was investigated based on the mechanism of phage-resistant cell conversion. Deletion of OmpC from the E. coli cells facilitated the emergence of cells resistant to SP21 phage. After 8 h of incubation, SP21-resistant cells appeared. By contrast, alteration of the lipopolysaccharide (LPS) profile facilitated cell resistance to SP22 phage, which was observed following a 6-h incubation. When a cocktail of phages SP21 and SP22 was used to infect E. coli O157:H7 cells, 30 h was required for the emergence of cells (R-C) resistant to both phages. The R-C cells carried almost the same outer membrane and LPS components as the wild-type cells. However, the reduced binding ability of both phages to R-C cells suggested disturbance of phage adsorption to the R-C surface. Even though R-C cells resistant to both phages appeared, this work shows that rational selection of phages has the potential to at least delay the emergence of phage resistance.     

Molecular typing of Salmonella weltevreden and Salmonella chincol by pulsed-field gel electrophoresis (PFGE) and enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR)

Genomic DNA of Salmonella weltevreden (10 isolates from poultry, two isolates each from raw vegetables and river water) and S. chincol (15 isolates from poultry) were characterized by pulsed-field gel electrophoresis (PFGE) and enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) analysis. These isolates originated from a single location in Kajang, Selangor. The results of the PFGE and ERIC-PCR were analysed and comparisons were made using GelCompar software. ERIC-PCR with primers ERIC1R and ERIC2 discriminated the S. weltevreden into five clusters and two single isolates and S. chincol into two clusters and two single isolates at a similarity level of 80%, respectively. PFGE produced a single cluster and eight single isolates for S. weltevreden, and one cluster and 11 single isolates for S. chincol at a similarity level of 80% after digestion with the restriction enzyme XbaI, respectively. These results demonstrate that both PFGE and ERIC-PCR are suitable tools for molecular typing of the isolates examined. This revised version was published online in July 2006 with corrections to the Cover Date.     

Large Variabilities in Host Strain Susceptibility and Phage Host Range Govern Interactions between Lytic Marine Phages and Their Flavobacterium Hosts

Phages are a main mortality factor for marine bacterioplankton and are thought to regulate bacterial community composition through host-specific infection and lysis. In the present study we demonstrate for a marine phage-host assemblage that interactions are complex and that specificity and efficiency of infection and lysis are highly variable among phages infectious to strains of the same bacterial species. Twenty-three Bacteroidetes strains and 46 phages from Swedish and Danish coastal waters were analyzed. Based on genotypic and phenotypic analyses, 21 of the isolates could be considered strains of Cellulophaga baltica (Flavobacteriaceae). Nevertheless, all bacterial strains showed unique phage susceptibility patterns and differed by up to 6 orders of magnitude in sensitivity to the same titer of phage. The isolated phages showed pronounced variations in genome size (8 to >242 kb) and host range (infecting 1 to 20 bacterial strains). Our data indicate that marine bacterioplankton are susceptible to multiple co-occurring phages and that sensitivity towards phage infection is strain specific and exists as a continuum between highly sensitive and resistant, implying an extremely complex web of phage-host interactions. Hence, effects of phages on bacterioplankton community composition and dynamics may go undetected in studies where strain identity is not resolvable, i.e., in studies based on the phylogenetic resolution provided by 16S rRNA gene or internal transcribed spacer sequences.     

Molecular characterization and module composition of P22-related Salmonella phage genomes.

Genomes of newly isolated Salmonella phages were analysed by comparison of their EcoRI restriction patterns and by hybridization. Characteristic hybridization probes from reference phages P22, ES18 and E. coli phage lambda were chosen. Four probes selected from the lysis region examined the dispersal of the lambdoid lysis genes. Other probes characterized were the replication genes and part of the structural genes. The complex immunity region was investigated by means of hybridization as well as biological tests. The results showed the relationship of the isolated phages to the P22 branch of the lambdoid phages and revealed their modular genome organization consisting of different proportions of P22-related sequences. DNA restriction patterns of phages released from Salmonella strains sampled in limited geographical areas were significantly less heterogeneous than those of phages released from the worldwide sampled SARA collection. The use of prophage restriction patterns as a tool for the typing of Salmonellae to support the epidemiologic classification of pathogenic strains is discussed.