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PubMed1975

Studies on a smooth phage resistant variant of Brucella abortus II. Mechanism of phage resistance.

The properties of a smooth phage resistant variant of Brucella abortus were studied in an attempt to determine the mechanism of phage resistance. This strain was fully capable of adsorbing phage but penetration, and hence replication, did not occur. No evidence of lysogeny or a phage carrier state could be obtained and gross chemical differences between the resistant strain and its phage susceptible parent were not detected. The phage resistant variant showed increased resistance to lysis from without, either by phage or by lysozyme, in the presence of chelating agents. It was concluded that resistance was the result of a modification in cell wall structure conferring resistance to lysozyme-like enzymes, thus preventing penetration of phage.

Adsorption↗
PubMed1976

[Effect of mutagens on RNA-containing phages and its infectious RNA. VII. Genetic nature of morphologic mutants of RNA-containing phage MS2].

Temperature-sensitive "leaky" mutants of phage MS2 having white dense ring around negative colonies are described. As these mutants are used for quantitative genetic studies, the white ring presents interest. Typical mutant 40 is used as a model for investigation. Light microscopy has shown, that cells from white ring zone have spore-like inclusions, which determine the characteristic structure of surrounding mutant negative colonies. Cytochemical reactions for the presence of glicogen, lipids, volutin, nuclear material and spores were negative. Electrone microscopy of negatively stained samples and ultrathin sections has revealed that cells from white ring zone, unlike phage-infected wild type cells, have two types of electron dense inclusions: 1) crystalline structures formed with great number of closely packed mature phage particles, and 2) large amorphic bodies. Electrone microscope-cytochemical data showed that inclusions remain intact under treatment of ultrathin sections of white zone ring with DNase and perchloric acid, while nuclear material was completely destroyed. Amorphic bodied were completely destructed after the treatment with RNase, while nuclear material and crystalline phage aggregated remained unchanged. Therefore, amorphic bodies consist of RNA, which has not been used to form virions. Single cycle of the development of mutant 40 at 37 degrees and 43 degrees C and under the temperature of incubation 37 degrees leads to 43 degrees C and 43 degrees leads to 37 degrees C in the course of intracellular reproduction is investigated. Influence of the phage on growth on infected culture is studied. The data obtained draw to a conclusion that the impaired function belongs to cystron protein of the phage membrane. As certain mutations in this cystrone of RNA-containing phage result in the depression of cystrone RNA polymerase, it is supposed that the formation of RNA containing bodies in infected cells, determining the formation of white rings in NA, together with cristalline aggregates of cells, is a result of mutation damage of cystrone protein of the phage MS2 membrane.

Coliphages↗
PubMed2026

Magnetite facilitates phage-bacteria interactions and phage-associated metabolic coordination for medium-chain fatty acid biosynthesis under ammonia stress.

Medium-chain fatty acid (MCFA) production from waste activated sludge (WAS) is considered to rely on the syntrophic interactions among distinct functional microorganisms. Whether phages represent an overlooked ecological component involved in MCFA biosynthesis remains unclear. MCFA production is often inhibited by high total ammonia nitrogen (TAN). Conductive materials have been proposed as effective strategies to enhance or recover MCFA production under TAN stress. Therefore, the inhibition-recovery system could provide a useful framework for investigating whether viral ecological responses are associated with MCFA-related metabolic processes. In this study, magnetite (Fe3O4) was selected as the recovery strategy of MCFA production under TAN stress. Results showed that a total of 3915 vOTUs and 118 metagenome-assembled genomes were recovered from the anaerobic bioreactors subjected to three conditions: Control (without TAN stress), TAN stress, and TAN stress with Fe3O4 presence. Under high TAN environment (∼5 g/L), MCFA production reduced by 49% (1.3 g COD/L) in comparison to that without high TAN stress. The ecological coupling between temperate phages and MCFA-related bacterial hosts was weakened, accompanied by substantial decreases in the abundance and transcriptional abundance of phage-associated auxiliary metabolic genes (AMGs) related to MCFA synthesis (e.g., fatty acid biosynthesis, acyl-chain metabolism). In the presence of Fe3O4 (i.e., 10 g/L), MCFA production was four and two times higher than those with and without high TAN stress. In addition to enhanced phage-bacteria ecological coupling and increased the abundance and transcription of AMGs related to MCFA formation, Fe3O4 increased the abundance and expressions of electron-transfer-related AMGs (e.g., cbb3-type cytochrome c oxidase, type IV pilus assembly genes) and QS-related LuxR-family and HTH-type regulators, indicating that such indirect pathway could be largely overlooked during MCFA synthesis. This is also the first-time reporting that phages could represent an ecological layer responsive to iron oxide. Analyses of publicly available metagenomes collected from MCFA-oriented anaerobic systems further confirmed that phages could be broadly associated with the metabolic processes involved in MCFA biosynthesis. Taken together, this study reveals that phages could serve as an overlooked ecological layer associated with MCFA metabolism and provide a viral-ecology perspective for understanding TAN inhibition and iron oxide-mediated recovery during WAS-to-MCFA bioconversion.

Ammonia inhibition↗
PubMed1978

Preliminary results of phage typing of coagulase-negative staphylococci with the set of typing phages of Pulverer and co-workers.

120 strains of Staphylococcus epidermidis isolated from clinical material were typed with the set of 16 test phages described by Pulverer et al. (1975). 58 (48%) strains were typable with RTD and additionally 12 strains with 100 times RTD. The pattern Ph10/U14/U16 was observed most common (36 strains), followed by the pattern U14/U16 (11 strains). One strain showed lysis reactions with all typing phages. With 61 strains phage U14 gave a lysis whereas phage U20 reacted only with 4 strains. The S. epidermidis phages were not specific for this species, at least some of these showed lysis reactions with S. aureus strains. The lysis pattern most often observed was Ph10/U14/U16. There was no correlation between the patterns given by the S. aureus test phages and those by the S. epidermidis phages.

Bacteriophage Typing↗
PubMed1976

Specialized transducing phages derived from phage P22 that carry the pro AB region of the host, Salmonella typhimurium: genetic evidence for their structure and mode of transduction.

Two independently isolated specialized transducing phages, P22pro-1 and P22pro-3, have been studied. Lysates of P22pro-1 contain a majority of transducing phages which can go through the lytic cycle only in mixed infection; these defective phages transduce by lysogenization in mixed infection and by substitution in single infection. A few of the transducing phages in P22pro-1 lysates appear to be non-defective, being able to form plaques and to transduce by lysogenization in single infection. Transduction by P22pro-3 lysates is effected by non-defective transducing phages, which transduce by lysogenization; these lysates also contain a majority of defective phages which do not co-operate in mixed infection. The P22pro-1 genome is thought to contain an insertion of bacterial DNA longer than the terminal repetition present in P22 wild type, so that at maturation a population of differently defective phages is produced. The exact structure of the P22pro-3 genome is open to conjecture, but it seems clear that the insertion of bacterial DNA is smaller than that in P22pro-1. Both P22pro-1 and P22pro-3 are defective in integration at ataA under non-selective conditions, although both integrate on medium that lacks proline.

Chromosome Mapping↗
PubMed2026

The value of a prophage-borne defense system in phage-phage competition.

Temperate phages that incorporate into their bacterial hosts' genomes often encode defense systems that protect their hosts from superinfection by unrelated phages. Yet the evolutionary value of such defenses to the phage remains unclear. We present a minimal theoretical framework to quantify the selective advantage of a prophage-borne defense system in competition between temperate phages infecting the same bacterial host. The model reveals regimes in which a "defensive phage" can invade and persist despite growth costs, regimes of bistability, and others in which all phage types coexist due to a rock-paper-scissors-like dynamic between defensive, non-defensive, and defense-loss variants. Because defense systems can be non-transitive, true rock-paper-scissors relations can lead to persistent oscillations. These results identify simple conditions under which phage-encoded defense systems are evolutionarily stable, providing testable predictions for the prevalence and maintenance of these systems in natural microbial communities.

Prophages↗
PubMed1978

[Phage-typing modifications induced by "in vitro" transfer of R plasmids. I.--Phage typing of Salmonella typhi (author's transl)].

The phage-typing modifications induced by transfer of antibiotic-resistance plasmids wre studied in two S. typhi Vi+ strains: n 2411 (phage-type A) and Ty2 (phage-type E1a). Forty-one R plasmids belonging to twenty-two incompatibility groups were investigated. Twenty-two plasmids were unable to produce any phage typing modifications. Among the groups of plasmids with phage-typing restriction capacity,four (I1, 10.B.O., N and W) groups were found heterogeneous with regard to this property and one (F1 group) caused significant modifications of the phage-types defined by the Vi phage-typing international system.

Anti-Bacterial Agents↗
PubMed1975

Staphylococcus aureus-strains of phage group II and their possible relation to animal staphylococci. 1. Antigenic properties, phage typing, and resistance to antibiotics.

Investigations carried out on 350 strains isolated from clinical materials and carriers revealed that among coagulase-positive staphylococci of phage group II two main serologic types predominated. The first (61,1%) was characterised by positive reaction with factor serum 263-2, the second (30,8%) one gave positive reaction with factor serum i1-2. The strains of type 263-2 contained as a rule polysaccharide A as well as polysaccharide 263, contrary to staphylococci belonging to type i1-2 which possessed polysaccharide A only. Both serologic types differed also epidemiologically, in phage typing and resistance to antibiotics. Staphylococci of serologic type 263-2 occurred in neraly all purulent lesions, they were more resistant to antibiotics than type i1-2 and prevailed in hospital environments. In phage-typing the differences between both serologic types were quantitative in character, with two being most outstanding, namely the prevalence of strains sensitive to phage 71 among staphylococci of serologic type 263-2 and of phage types 3A, and 3C as well as 3A, 3B, 3C among staphylococci of serologic type i1-2. There was no correlation between the susceptibility of strains to phage 71 and their virulence, the latter seemed to be connected with serologic type.

Anti-Bacterial Agents↗
PubMed1976

[Phage typing of cholera vibrios with different sets of phages].

A comparative study was made of two sets of Mukerjee and Drozhevkina-Arutyunova's bacteriophages in typing 514 strains of the El Tor vibrios and 45 strains of clasic biotype. It was shown that the Mukerjee or Drozhevkina-Arutyunova's phages could be used for the typing of cholera vibrios. The phages of the latter set prove to detect more phage types (18 against 11); they determine both the phage type and the biotype at the same time. The typing of cholera vibrios of both biotypes is possible, and the percentage of nontyping strains left is comparatively low (5.2 against 23.5 after Mukerjee). A table of the phage correspondence was made; it permits to obtain comparable data in using any set of the typing phages.

Bacteriophage Typing↗
PubMedDate not supplied

[Resistance to phage MS2 induced in E. coli by infection with that phage].

A sensitive cell of E. coli AB 259 Hfr 3000 infected with RNA-containing phage MS2 produces phages and simultaneously continues to divide showing a segregation of sensitive cells which maintain new cycles of the infection. Phage multiplication in the sensitive cell induces phage-resistant forms in the progeny of this cell. The described phenomenon is not caused by selection of pre-existing F--cells, but may result from a direct interaction of phage products with the episomal DNA coding proteins for F-pili. The elucidation of the mechanisms of this phenomenon may pave the way to studies of the DNA- and RNA- containing genome interaction within the cell, the mechanisms of persistent infection and the causes of different degrees in phage virulence.

Coliphages↗
PubMedDate not supplied

[Phage T4 partial diploidy obtained with the method of DNA interrupted injection. I. Analysis of the genetic structure and phage progeny reproduction process].

Phage T4 chromosome fragmentation is shown to take place when DNA injection is interrupted, a fragment length being strictly controlled by the interval from the moment of adsorbtion till the moment of an interruption. Populations of the bacteria cells infected by the phage T4 partial diploids are produced with the method of DNA interrupted injection. In the population a merodiploid involves some phage T4 amber mutant and a phage "wild" type chromosome fragment of the size controlled. To construct merodiploids the amber mutant in gene 43 and the mutant in gene 32 with the higher and the lower recombination frequency, accordingly, are used. Every merodiploid which is the heterozygote by one of these genes or which is the heterozygote by the late genes is determined to reproduce mixed phage progeny. Both the mean of the burst and the parent genotypes ratio in progeny either in the E. coli CR-63 cells or in the E. coli B depend on neither the heterozygote genetic structure nor the diploid region size. The results obtained conclude that phage genes express their function in the small fragments and the fragment recombination with the mutant partner whole chromosome follows their autonomous replication.

Chromosomes↗
PubMed1975

Modification of Escherichia coli membranes in the prereplicative phase of phage T4 infection. Specificity of association and quantitation of bound phage proteins.

Reinfection of Escherichia coli with the bacterial virus T4 causes modifications of the properties of the host cell envelope during the preeplicative phase of the lytic cycle. These changes include altered densities cell enveloped and their subfractions, morphological modifications of membrane vesicles, and association of newly synthesized proteins with the host cell envelope. Polypeptide analysis by high resolution electrophoresis on polyacrylamide slab gels in dodecyl sulfate revealed that most of some 30 prereplicative phage-coded polypeptides are attached to this structure. Different means of cell disruption and selective extraction procedures, such as variations of ionic strength, removal of divalent cations, and the addition of chaotropic agents or detergents were used to study the characteristics of these attachments. Many proteins appeared to be artifactually absorbed or weakly bound to the envelope, Separation of cell walls from plasma membranes showed that all of the tightly bound proteins were associated withthe cell membrane fraction. The partitioning of phage proteins between the different fractions was monitored using 12 polypeptides which were identified as products of distinct phage genes. Of these, 8 were eliminated as potential membrane markers. Four polypeptides, the products of genes rIIA, rIIB, 39, and 52 were operationally defined as membrane proteins. The number of molecules of the 12 identified phage gene products, synthesized during a single lytic cycle, was determined. The results allowed the estimation of the concentration in the membrane of those proteins which were found to be quantitatively associated with that structure. Association of phage proteins with the cell envelope was found to be unaffected by mutations in any of the identified phage polypeptides.

Amino Acids↗
PubMed1978

Phage typing of nontypable isolates of Staphylococcus aureus using the new phages 94, 95, and 96 and the heat-shock treatment.

Of 2,144 isolates of Staphylococcus aureus (from human beings in New York State) received in the authors' laboratory during 1975, 992 (46.,%) were nontypable with the use of the international basic set of bacteriophages. These 2,144 isolates were retested with the use of the recently added phages 94, 95, and 96. Nearly 60% of the isolates were lysed with one or more of these three phages, including 64% of those previously nontypable, thus reducing the percentage of nontypables to 16.5%. One hundred randomly selected isolates that remained nontypable were heat-shocked and retested with the revised basic set of phages, which includes the three new phages. Forty-three isolates were typable with the use of this procedure; all but one of these produced the exact same phage pattern in at least three of five trials.

Bacteriophage Typing↗
PubMed1979

Salmonella phage glycanases: substrate specificity of the phage P22 endo-rhamnosidase.

Interaction between phage P22 and phenol-water extracted lipopolysaccharides from sensitive Salmonella bacteria belonging to serogroups A, B and Di results in hydrolysis of the alpha-L-rhamnosyl linkages within the tetrasaccharide repeating unit of the O-antigenic polysaccharide chain. These O-antigens have identical structures except for the nature of the 3,6-dideoxy-hexosyl group linked to O-3 of the D-mannosyl residue. Removal of the dideoxysugar, or periodate oxidation followed by borohydride reduction of the L-rhamnosyl residue made the O chain resistant to the endo-rhamnosidase. Substitution of the D-galactosyl residue at O-4, but not at O-6, with an alpha-D-glucosyl group was compatible with hydrolysis. A number of Klebsiella pneumoniae and Shigella flexneri lipo- or capsular polysaccharides containing chain L-rhamnosyl residues were tested but none was sensitive to the P22 endo-rhamnosidase. The substrate specificity of the endo-rhamnosidase parallels the lytic specificity of the phage which suggests that the initial step in phage P22 infection is a P22 tail enzyme O-antigen substrate interaction. The main product of the hydrolysate was octa-, dodeca- and hexadecasaccharides. Treatment of phage FO resistant smooth strains of S. typhimurium with P22 tails removed O polysaccharide chains and made previously 'hidden' FO receptors accessible to the phage.

Glycoside Hydrolases↗
PubMed1977

Phage-typing of Salmonella weltevreden based on lysogeny. I. The phage-typing system.

All of the 1070 strains of Salmonella weltevreden tested were lysogenic, temperate phage being demonstrable in 83.9% of the strains in broth culture, 6.4% in mixed culture and the remaining 9.7% after induction by UV-irradiation and MC treatment. A phage-typing scheme for Salmonella weltevreden based on the host range of temperate phages was developed. The strains could be classified into eight groups by means of the host range of their temperate phages on six indicator strains and a ninth group, the symbiotic phages of which failed to lyse any of the indicator strains.

Bacteriophage Typing↗
PubMed1976

On the lack of host-cell reactivation of UV-irradiated phage T5. I. Interference of T5 infection with the host-cell reactivation of phage T1.

UV-irradiated phage T5, in contrast to T1, T3 and T7, fail to display host-cell reactivation (HCR) when infecting excision-repair proficient Escherichia coli cells. Possible causes of this lack of HCR (which T5 shares with the T-even phages) have been investigated by studying HCR of T1 under conditions of superinfection by T5. Repair-proficient B/r cells were infected at low multiplicity with UV-irradiated phage T1 in the presence of 1.8 mg/ml caffeine and were superinfected after 15 min with heavily UV-irradiated T5 amber mutants at highly multiplicity. The caffeine, which is later diluted out, prevents any T1 repair prior to T5 superinfection, and UV (254 nm) irradiation of T5 with 144 J/m2 reduces the ability of this phage to exclude T1, thus permitting a reasonable fraction of the mixedly infected complexes to produce T1 progeny. Under these conditions, T5 superinfection causes loss of HCR in about 90% of the T1-producing complexes. Superinfection with unirradiated T5 likewise inhibits HCR of T1, but superinfection with irradiated T3 (a host-cell reactivable phage) does not. This indicates that the observed HCR inhibition of T1 results from T5 infection rather than from competition of irradiated foreign DNA for the excision-repair enzymes of the bacterial host. Employment of appropriate T5 amber mutants has shown that "first-step transfer" (FST) of T5 DNA (involving only 8% of the T5 genome) is sufficient for HCR inhibition, but that transfer of the remainder DNA in addition inhibits a previously described minor T1 recovery process. HCR inhibition of T1, and thus presumably lack of HCR in T5 itself, is ascribed to a substance which is produced either post infection by a gene located in the FST segment of the T5 genome, or which is transferred from extracellular T5 together with the FST DNA.

Coliphages↗
PubMed1977

[Site-specific recombination between phages lambda and phi 81 and integration of hybrid phages lambda-phi 81].

Dependence of the formation frequency of hybrid phages immlambdahphi81 and immphi81hlambda in recombination between phages lambda and phi81 from int-function of both phages is studied. Phages with hybrid att-sites (Plambda OP'phi81 and Pphi81OP'lambda) are isolated and the efficiency of integration of these phages into bacterial chromosome is determined.

Chromosomes, Bacterial↗
PubMed1978

The activity of ant product of the Salmonella phage P22 against the closely related but heteroimmune phage L.

P22 mutants defective in the early gene 24 are complemented by phage L in mixed infection. P22 12- and P22 23- mutants are not complemented by phage L. Gene function 24 of an L prophage is turned on by a superinfecting P22 24- mutant and complements the missing function of the defective P22 phage. Since this transactivation of prophage gene 24 depends on a functional gene ant in the superinfecting P22 mutant, it indicates derepression for leftward directed gene expression in prophage L. On the contrary neither the rightward directed expression of gene 12 nor of gene 23 in prophage L. can be turned on by superinfecting P22 24- 12- or P22 24- 23- mutants (and also not by P22 12- and P22 23-) to a degree sufficient for complementation of simultaneously superinfecting L virB 12- or L virB 23- mutants. The failure to detect release of repression for rightward directed gene expression of prophage L corresponds to the earlier observation (Prell, 1975) that P22 superinfecting L lysogens cannot release replication inhibition for simultaneously infecting phage L. The results are discussed with respect to the mechanism underlying the different action of P22 antirepressor in L and in P22 lysogens.

DNA Replication↗