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Coliphage BA14: a new relative of phage T7.

Coliphage BA14 was isolated from sewage and shown to be related to phages T7 and T3. It is similar to T3 in that it directs the synthesis of an S-adenosyl-methionine-cleaving enzyme (SAMase) early upon infection. However, it differs from all other known T7-related coliphages by the inability of its RNA polymerase (gene 1 product) to transcribe T7 DNA or T3 DNA. BA14, T7 and T3 also show marked differences in autoradiographic patterns of their gel-electrophoretically separated 35S-labelled intracellular phage proteins, restriction endonuclease HpaI cleavage patterns of their DNAs, and serological specificities of their infectious particles. Other distinctive features became apparent upon simultaneous mixed infection with BA14 and T7 or T3: inability of BA14 to produce genetic recombinants with either T7 or T3; lack of functional complementation between amber mutants of BA14 and T7 or T3; mutual exclusion and depression of the burst size of the mixedly infected cells.

Antigens, Viral↗

The host-dependent restriction of growth of an RNA coliphage FI.

Phage FIC is a spontaneous host-dependent mutant of phage FI which is classified into the fourth group of RNA Escherichia coli phages (RNA coliphages). The mutant phage (FIC) grows normally in E. coli strain Q13 (permissive host), but poorly in strain A/lambda (non-permissive host) (9). Attempts to elucidate the regulatory mechanism of growth of the mutant phage in the non-permissive host revealed the following: (a) growth of the mutant phage was specifically restricted in E. coli strains that have certain suppressor genes for amber mutation; (b) the mutant phage RNA (FIC-RNA) could not produce progeny in the spheroplasts of the non-permissive host; (c) adsorption of the mutant phage to, and penetration of the mutant phage RNA into, the non-permissive host were normal; and (d) biosynthesis of the phage-specific late protein and RNA did not occur in the non-permissive host. Based on these results we conclude that phage FIC is a spontaneous azure-type mutant of the fourth group of RNA coliphage FI.

Adsorption↗

Coliphage HK243: biological and physicochemical characteristics.

Coliphage HK243 can form plaques on Escherichia coli C and K-12, but not B. The plaques are 1-2 mm in diameter and are opaque areas which clear upon exposure to chloroform vapor. During one-step growth, the eclipse and the latent periods are 20 and 30 min, respectively. Phage-infected cells continue to produce cell-free plaque-forming units for as long as 80 min after the end of the latent period, although at high multiplicities of infection (MOI) most cells lyse. No lysogenic bacteria have been found among survivors, so HK243 is considered a virulent phage. Some of the cells surviving a high MOI challenge are maltose negative and resistant to both HK243 and coliphage lambda. This fact has made possible the isolation of lambda-resistant mutants of lambda-lysogens. However, no serological cross-reaction between the phages lambda and HK243 has been detected. Genetic data involving three essential loci and a locus controlling plaque morphology suggest a circular linkage map. The virions are tadpole-shaped with an icosahedral head 68 nm long which is attached to a flexible tail 131 nm long. The phage has a linear, duplex DNA genome of molecular weight approximately 44 x 10(6) and a base composition of 33% adenine, 31% thymine, 16% guanine, and 20% cytosine.

Coliphages↗

Distribution of RNA coliphages in Senegal, Ghana, and Madagascar.

The distribution patterns of RNA coliphages (phages) in Senegal, Ghana, and Madagascar were investigated by collecting sewage samples from domestic drainage in November, 1980. In Senegal, among 65 sewage samples collected mainly from Dakar and its vicinity, 14 (22%) contained RNA phages (16 strains). By serological analysis, 13 of 16 strains were found to belong to group III. This is consistent with the distribution pattern of RNA coliphages in tropical and subtropical regions of Asia. In Ghana, however, among 106 samples collected from Accra, Suhum, and their vicinities, only seven (7%) contained RNA phages (seven strains) (groups I, II, and III [1:3:3]). In Madagascar, among 124 samples collected from Antananarivo, Moramanga, and their vicinities, seven (6%) contained RNA phages (seven strains) (groups I, II, III, and IV [1:1:1:4]). In spite of the low isolation frequency, it can be said that Madagascar appears to have a unique distribution pattern (abundance of group IV phages) which differs from that of any other countries we have examined. The generality of the distribution pattern of RNA phages in the tropical region (abundance of group III phages) was thus verified at least in Senegal.

Coliphages↗

Detection of coliphages and enteroviruses in sewage and aerosol from an activated sludge wastewater treatment plant.

Coliphages and enteroviruses were monitored over 12 months in sewage and air adjacent to an activated sludge plant. Both showed temporal variation but the mean count of phages in enterovirus-positive samples was not significantly different from that in enterovirus-negative samples. Hence coliphages are not necessarily a good indicator of enteroviruses in sewage and aerosols.

Aerosols↗

Role of lime treatment in the removal of bacteria, enteric viruses, and coliphages in a wastewater reclamation plant.

Lime flocculation/sedimentation in the first process unit of a 4,500-m3/day wastewater reclamation plant reduced numbers of microorganisms extensively when operated at pH 11.2. The efficiency was much less at lower pH values, and some bacteria even multiplied at pH 9.6. Data on reduction in the number of microorganisms in the lime treatment and subsequent units indicate that inactivation by hydroxide alkalinity plays an important role in the efficiency of lime treatment. Reductions in the numbers of enteric viruses were higher than those of coliphages, enterococci, and total plate and coliform bacteria, which indicate that lime treatment can be monitored by means of coliphage and conventional bacteriological tests. This paper illustrates the valuable role of high-pH lime treatment in reducing the load of pathogenic microorganisms on subsequent units, including ultimate disinfection processes, which is important in the multiple safety barrier concept of wastewater reclamation processes.

Bacteria↗

Distribution of ribonucleic acid coliphages in south and east Asia.

We investigated the distribution of ribonucleic acid (RNA) coliphages in the Philippines, Singapore, Indonesia, India, and Thailand by collecting sewage samples from domestic drainage in November 1976. Of the 221 samples collected from domestic drainage, 50 contained RNA phages (52 strains). By serological analysis, 46 of the 52 strains were found to belong to group III. It can thus be said that the most prevalent RNA phages in Southeast Asia (at least, in the Philippines, Singapore, and Indonesia) were group III phages. Investigations of sewage samples collected from domestic drainage in Japan indicate that the most prevalent RNA phages in mainland Japan (north of Kyushu) are group II phages, whereas group III phages are predominant in the southern part of Japan (south of Amamiohshima Island). We therefore propose a borderline between Kyushu and Amamiohshima Island for the geographical distribution of RNA coliphages in the domestic drainage of South and East Asia. Moreover, one strain (ID2) was inactivated to some extent with the antisera of four groups of RNA phages. This is thought to be significant from the evolutionary viewpoint.

Coliphages↗

Toxicity of zinc to fungi, bacteria, and coliphages: influence of chloride ions.

A 10 mM concentration of Zn2+ decreased the survival of Escherichia coli; enhanced the survival of Bacillus cereus; did not significantly affect the survival of Pseudomonas aeruginosa, Norcardia corallina, and T1, T7, P1, and phi80 coliphages; completely inhibited mycelial growth of Rhizoctonia solani; and reduced mycelial growth of Fusarium solani, Cunninghamella echinulata, Aspergillus niger, and Trichoderma viride. The toxicity of zinc to the fungi, bacteria, and coliphages was unaffected, lessened, or increased by the addition of high concentrations of NaCl. The increased toxicity of zinc in the presence of high concentrations of NaCl was not a result of a synergistic interaction between Zn2+ and elevated osmotic pressures but of the formation of complex anionic ZnCl species that exerted greater toxicities than did cationic Zn2+. Conversely, the decrease in zinc toxicity with increasing concentrations of NaCl probably reflected the decrease in the levels of Zn2+ due to the formation of Zn-Cl species, which was less inhibitory to these microbes than was Zn2+. A. niger tolerated higher concentrations of zinc in the presence of NaCl at 37 than at 25 degrees C.

Bacteria↗

Temperate coliphages: classification and correlation with habitats.

Temperate coliphages were recovered from sewage, mammalian feces, and lysogenic strains of Escherichia coli. A total of 32 phages of independent origin were divided into six groups by applying the criteria of host range, antigenic homology, and the ultraviolet inducibility of the prophage. The demonstration of genetic interactions in some cases has confirmed the classification scheme. Nine phages were assigned to the P2 family and 19 to the lambda family. The remaining four isolates may represent some novel phylogenetic types. Phages recovered from the lysogenic strains of E. coli were all found to be P2 related, whereas a majority of the phages recovered as cell-free plaque-forming units were assignable to the lambda family. It is proposed that the biological attributes of the phages belonging to the two principal families are reflected in the distribution patterns observed. The virions of phage HK256 show multiple tail fibers and may thus represent a "new" virion form among the temperate coliphages.

Bacteriophage lambda↗

Survival of F-specific RNA coliphage, feline calicivirus, and Escherichia coli in water: a comparative study.

The relationship between the survival of enteric viral pathogens and their indicators (coliform bacteria and coliphages) is not well understood. We compared the survival rates of feline calicivirus (FCV), Escherichia coli, and a male-specific RNA coliphage MS2 at 4, 25, and 37 degrees C for up to 28 days in dechlorinated water. The survival rates of E. coli and FCV, a surrogate of noroviruses (NV), had a high degree of correlation at 4 and 25 degrees C, while MS2 phage survived significantly longer (P < 0.05) at these two temperatures. At 37 degrees C, the survival rates for all three organisms were highly correlated. Decimal reduction values indicating the number of days needed for 90% reduction in titer (D values) decreased for all three organisms as storage temperatures increased. FCV had the shortest D value among all three organisms at all temperatures investigated. These findings indicate that F-specific RNA phages may be useful indicators of NV in the environment.

Calicivirus, Feline↗

DNA replication studies with coliphage 186: the involvement of the Escherichia coli DnaA protein in 186 replication is indirect.

The inability of coliphage 186 to infect productively a dnaA(Ts) mutant at a restrictive temperature was confirmed. However, the requirement by 186 for DnaA is indirect, since 186 can successfully infect suppressed dnaA (null) strains. The block to 186 infection of a dnaA(Ts) strain at a restrictive temperature is at the level of replication but incompletely so, since some 20% of the phage specific replication seen with infection of a dnaA+ host does occur. A mutant screen, to isolate host mutants blocked in 186-specific replication but not in the replication of the close relative coliphage P2, which has no DnaA requirement, yielded a mutant whose locus we mapped to the rep gene. A 186 mutant able to infect this rep mutant was isolated, and the mutation was located in the phage replication initiation endonuclease gene A, suggesting direct interaction between the Rep helicase and phage endonuclease during replication. DNA sequencing indicated a glutamic acid-to-valine change at residue 155 of the 694-residue product of gene A. In the discussion, we speculate that the indirect need of DnaA function is at the level of lagging-strand synthesis in the rolling circle replication of 186.

Bacterial Proteins↗

Inactivation of the T-even coliphages by pyrophosphate.

Van Vunakis, Helen (Brandeis University, Waltham, Mass.) and Roger M. Herriott. Inactivation of the T-even coliphages by pyrophosphate. J. Bacteriol. 83:590-596. 1962.-Pyrophosphate liberates the deoxyribonucleic acid from the T-even coliphages. The reaction is dependent on pH, temperature, time, and ionic environment. The reaction can be inhibited by magnesium, putrescine, spermidine, and neutral molecules above a certain molecular weight. Possible mechanisms of the reaction are discussed.

Bacteriophages↗

Growth of coliphage T7 in Salmonella typhimurium.

A mutant of Salmonella typhimurium was found to be sensitive to killing by coliphage T7 because of an alteration in its surface properties. However, the infections were abortive and studies with (32)P-labeled T7 grown in Escherichia coli B (T7.B) indicated that the phage DNA was restricted by S. typhimurium. When a mutant T7 which survived the restriction and produced plaques on Salmonella (T7.S) was passed through one cycle of growth in E. coli B, its ability to grow in Salmonella was lost, indicating that host-controlled restriction and modification are operative in this system. Restrictionless S. typhimurium mutants were isolated that permit the growth of not only T7.S but also T7.B and coliphage T3. The physiology of T7 production in the restrictionless host is nearly identical to that in Escherichia coli.

Alkanesulfonates↗

Prophage induction of noninducible coliphage 186.

Coliphage 186 has been regarded as a member of the noninducible group of coliphages. Evidence that prophage 186 is induced by ultraviolet irradiation or by treatment with nalidixic acid or mitomycin C is now presented. The phage yields were similar to those from lysogens of the inducible phage lambda, and the induction required a recA(+) host. A noninducible mutant of 186 was isolated from its heat-inducible derivative, 186cIts, that was no longer inducible by ultraviolet irradiation but remained heat inducible. That zygotic induction of 186 after transfer from a lysogenic male to a non-lysogenic recipient did not occur is indicated by the following findings: (i) there was only a slight increase in phage titer; (ii) similar levels of recombinants were obtained for markers adjacent or distal to the phage integration site, whether the recipient was lysogenic or not, and there was no effect on the gradient of marker transfer; (iii) lysogenic recombinants were readily found and the co-transfer of 186 with adjacent markers was the same to lysogenic or non-lysogenic recipients. Thus, 186 formed an inducible prophage that did not display zygotic induction. Nevertheless, it shared many properties with the noninducible phage P2 as outlined in the discussion.

Coliphages↗

Coliphages and enteric viruses in the particulate phase of river water.

The present study was undertaken to determine if indigenous enteric viruses and coliphages are free or associated with suspended particulate matter in natural waters. River water was filtered on filters of decreasing porosities (100-0.25 micron) that were pretreated with detergent to eliminate viral adsorption while retaining particulates. This filtered water was refiltered in virus-adsorbing conditions to retain free viruses. The virus-adsorbing filter retained most of the enteric viruses (77.4%) and coliphages (65.8%), which indicated that these viruses were probably free or associated with particles with a diameter of less than 0.25 micron. These observations are important because in water treatment plants small particulates are often the most difficult to eliminate.

Adsorption↗

Concentration of Giardia lamblia cysts, Legionella pneumophila, Clostridium perfringens, human enteric viruses, and coliphages from large volumes of drinking water, using a single filtration.

Poliovirus, coliphages, Giardia lamblia cysts, Clostridium perfringens spores, and Legionella pneumophila were concentrated simultaneously in a single pass by sequential filtration of large volumes of drinking water through 3- and 1-micron wound electronegative fiberglass cartridge filters (25.4 cm). Filtration was performed under acidic conditions (pH 3.5) in the presence of 0.001 M aluminum chloride to enhance adsorption. Elution of all the microorganisms entrapped or adsorbed to the filters was obtained by a slow backwash elution with a 1.5% beef extract solution, pH 9.75, containing 0.5% Tween 80. Tween 80 was shown to enhance recovery of the bacteriophages, bacteria, and parasites. Giardia cysts were efficiently eluted (71%) and could be reconcentrated by low-speed centrifugation and purified by sucrose density gradient flotation at a final recovery of 52%. Legionella pneumophila cells were eluted at 64% and were further concentrated by low-speed centrifugation at an overall recovery of 55%. C. perfringens spores and coliphages were eluted at efficiencies of 82 and 86%, respectively, and reconcentrated with minimal loss by a detergent - protein flotation method. Poliovirus was eluted at 93% and reconcentrated at 78% efficiency by organic flocculation.

Animals↗

Effect of the method of preparing monochloramine upon inactivation of MS2 coliphage, Escherichia coli, and Klebsiella pneumoniae.

Monochloramine prepared in situ by first adding chlorine to a suspension of microorganisms, followed by subsequent addition of ammonia, inactivated the MS2 coliphage more rapidly than did exposure of phage to monochloramine prepared either by adding chlorine to ammonia or by adding chlorine and ammonia simultaneously. The rapid viral inactivation was apparently due to the exposure of MS2 to free chlorine before the addition of ammonia. The average 99% CT value of MS2 when exposed to free chlorine was 1.3 and 1.1 at 5 and 15 degrees C, respectively. The average 99% CT values of MS2 briefly exposed to the combined action of free chlorine followed by the addition of ammonia to form monochloramine in situ were 19.3 and 1.5 at 5 and 15 degrees C, respectively. No 99% CT values were calculated for the inactivation of MS2 with preformed monochloramine because less than 1 log (90%) of inactivation occurred during a 4-h contact time. Inactivation of MS2 by monochloramine was more rapid at 15 than at 5 degrees C and when the chlorine to nitrogen weight ratio was 5:1 compared with 3:1. Monochloramine was a more efficient inactivating agent for the coliforms Escherichia coli and Klebsiella pneumoniae than it was for the MS2 coliphage.

Chloramines↗