Bacteriophage Ecology Group
Reference Abstracts (1989)
Dedicated to the ecology and evolutionary biology of the parasites of unicellular organisms (UOPs)
© Stephen T. Abedon
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© Phage et al. last updated on Wednesday, December 26, 2001
  1. Selection for bacteriophage latent period length by bacterial density: A theoretical examination. Abedon, S.T. (1989). Microb. Ecol. 18:79-88. In bacteriophage (phage), rapid and efficient intracellular progeny production is of obvious benefit. A short latent period is not. All else being equal, a longer latent period utilizes host cell resources more completely. Using established parameters of phage growth, a simulation of three successive phage lysis cycles is presented. I have found that high, but not low, host cell densities can select for short latent periods. This results from phage with short latent periods more rapidly establishing multiple parallel infections at high host cell concentrations, whereas phage with long latent periods are restricted to growth within a single cell over the same period. This implies that phage with short latent periods habitually grow in environments that are rich in host cells. [TOP OF PAGE]

  2. ??? Alippi, A.M. (1989). Microbiologia 5:35-43. [this is cited in Lederberg, 1996 (PNAS 93:3167-3168): "There have been suggestions that the synecology of bacteriophage and bacterial pathogens may account for fluctuations in their outbreaks (ref)"]. [TOP OF PAGE]

  3. Bacteriophage transport in sandy soil and fractured tuff. Bales, R.C., Gerba, C.P., Grondin, G.H., Jensen, S.L. (1989). Appl. Environ. Microbiol. 55:2061-2067. [TOP OF PAGE]

  4. Sequence counter-selection in cyanophage. Bancroft, I., Smith, R.J. (1989). p. 316 In Rogers, L.J. and Gallon, J.R. (eds.), BIOCHEMISTRY OF THE ALGAE AND CYANOBACTERIA. An analysis of the cleavage of native and cloned DNA of five cyanophage which infect Anabaena 7120 by 34 restriction endonucleases provided evidence of sequence counter-selection similar to that present in T sub(7). One group are isoschizomers of Anabaena) 7120 endogenous restriction endonucleases. Another group contain the subsequence GATC; and a third group contain dicytosine residues. The fourth group have no common sequence structure and may represent isoschizomers of restriction endonucleases present in the host range of the five cyanophage. Cyanophages AN-23, AN-13, and A-4L do not tolerate sequence methylation. A-1L and AN-10 tolerate adenine methylation, but differ in their tolerance of cytosine methylation. AN-10 appears able to prevent cytosine methylation by host enzymes. AN-10 and A-1L are closely related. Comparison of their restriction maps shows that counter-selection of some Hae III sites in AN-10 has occurred since divergence of the phage from their common ancestor. [TOP OF PAGE]

  5. The virulence characteristics of strains of Escherichia coli isolated from cases of bovine mastitis in England and Wales. Barrow, P.A., Hill, A.W. (1989). Veterinary Microbiology 20:35-48. Of 470 Gram-negative facultative anaerobes isolated from cases of bovine mastitis in England and Wales, 422 were identified as Escherichia coli. The characteristics of 237 of these were investigated. Guinea-pig red cell haemagglutinins were possessed by 86% of strains and 12% were resistant to D-mannose. None of the strains tested invaded Vero cells. Haemolysin, Vero toxin and enterotoxin were produced by 5, 0.5 and 1% of strains, respectively. Twenty-two percent were resistant to one or more antibiotics and 4% to sodium arsenate. Transfer ability was possessed by 41% and lysogenic phage by 27% of strains; 62% possessed either one or the other and 12% possessed both. Colicin production was detected in 18% of strains; 5% produced Colicin V. Ninety-nine percent of strains were serum-resistant, while only 6% were able to grow well in bovine serum. A microscopically visible capsule was seen in 75% of strains. All strains possessed at least one of the potential virulence factors or markers studied. Several strains which possessed one characteristic only (mannose-sensitive haemagglutination or serum resistance), possessed one or more large molecular weight plasmids. None of the strains was particularly virulent for chickens following intramuscular inoculation. Of the strains which possessed one virulence characteristic, only those which were serum-resistant were re-isolated from expressed milk following intramammary inoculation of lactating cows. [TOP OF PAGE]

  6. Economies of scale. The smallest of the small swim in unsuspected numbers. Beardsley, T.M. (1989). Scientific American November:22-22. So, naturalists observe, a flea/Hath smaller fleas that on him prey;/And these have smaller still to bite 'em;/And so proceeed ad infinitun. --Jonathan Swift. Swift's verse might not be literally true, but recent findings suggest he was closer to the truth than he probably realized. The majority of aquatic microorganisms were once thought to be nanoplankton, tiny organisms measuring no mre than 10 or 20 micrometers in diameter. Then picoplankton, organisms less than two micrometers across, were found to be far more numerous than had been realized. Now it seems that the world's water is teeming with life's smallest forms: viruses no bigger than .2 micrometer in diameter. [TOP OF PAGE]

  7. Bacteriophage introns: parasites within parasites? Belfort, M. (1989). Trends in Genetics 5:209-213. Several bacteriophage introns carry the information necessary to perform a complex series of RNA as well as DNA rearrangements. The RNA splicing reactions allow expression of the intron-containing genes, whereas recombination at the DNA level results in the mobility and invasive potential of the introns. The RNA and DNA transactions are temporally regulated to permit timely splicing early in infection, and expression of intron-encoded DNA endonucleases involved in mobility at late times. This is achieved through efficient use of the viral regulatory machinery, ensuring the perpetuation of the 'parasitic' introns. [TOP OF PAGE]

  8. Isolation and characterization of Klebsiella pneumoniae unencapsulated mutants. Benedi, V.J., Ciurana, B., Tomas, J.M. (1989). Journal of Clinical Microbiology 27:82-87. Klebsiella pneumoniae mutants were obtained after UV irradiation and negative selection with anticapsular serum. Unencapsulation, rather than expression of a structurally altered capsule, was found in the mutants. The mutant strains showed no alterations in their outer membrane proteins and lipopolysaccharide, and a great similarity with the wild type in the properties tested (serum resistance, antimicrobial sensitivity, and lipopolysaccharide-specific bacteriophage sensitivity), with the exception of a higher cell surface hydrophobicity and resistance to bacteriophage FC3-9. [TOP OF PAGE]

  9. High abundance of viruses found in aquatic environments. Bergh, O., Børsheim, K.Y., Bratbak, G., Heldal, M. (1989). Nature 340:467-468. [TOP OF PAGE]

  10. Isolation and characterization of a Pseudomonas aeruginosa bacteriophage with a very limited host range. Bigby, D., Kropinski, A.M.B. (1989). Can. J. Micribiol. 35:630-635. [TOP OF PAGE]

  11. Viruses in Water Systems: Detection and Identification. Block, J.C., Schwartzbrod, L. (1989). VCH, New York.[TOP OF PAGE]

  12. FRNA bacteriophages as indicators of fecal pollution in an estuarine environment. Boyd, D.M., Kator, H., Rhodes, M. (1989). Williamsburg, VA (USA). #1990 Annu. Meet. of the National Shellfisheries Association. 1900.A number of alternate microbial indicators of fecal pollution in receiving waters have been proposed. One of these, the male-specific coliphage-containing ribonucleic acid (FRNA), has not been evaluated in shellfish growing waters. The occurrence of FRNA phage in point source impacted water samples may be a better measure of pathogenic viral persistence than traditional coliform indicators. Accordingly, water and sediment samples were obtained from a tidal creek of the Ware River, Virginia, which receives treated effluent from a sewage treatment plant. Samples were assayed for male-specific phage using a method designed to enumerate FRNA phage lytic for Escherichia coli . Samples were collected along a salinity gradient over a 6 month period to evaluate the seasonal and spatial distribution of FRNA phage. Fecal coliform densities and selected physical/chemical parameters were also measured and compared with phage titer and occurrence. These results and the applicability of this assay method as an indicator of water quality in an estuarine watercourse impacted by a point source of sewage pollution are discussed. [TOP OF PAGE]

  13. Phage and fluorescent dyes tracing in karst. Bricelj, M., Krivic, P., Zupan, M. (1989). International Geological Congress, Abstracts--Congres Geologique Internationale, Resumes 28:199-200. [TOP OF PAGE]

  14. Role of the antilysozyme activity of Shigella flexneri as a factor controlling the degree of their resistance to the lytic action of bacteriophages. Bukharin, O.V., Deryabin, D.G. (1989). Zhurnal Mikrobiologii, Epidemiologii i Immunobiologii 16-19. The level of the antilysozyme activity of S. flexneri in ensuring the high level of their phage resistance has been studied. The realization of the phage protective effect of S. flexneri in ensuring the high level of their phage resistance has been studied. The realization of the phage protective effect of antilysozyme activity has been noted to occur due to disturbances in the lysis of infectd bacteria by phage-synthesized lysozyme- like enzyme. The direct relationship between the level of the lysozyme production of bacteriophages and their capacity for overcoming the antilysozyme protection of the host bacterium has been shown. [TOP OF PAGE]

  15. Cloning and expression in Escherichia coli of the genes of the arginine deiminase system of Streptococcus sanguis NCTC 10904. Burne, R.A., Parsons, D.T., Marquis, R.E. (1989). Infect. Immun. 57:3540-3548. The common oral bacterium Streptococcus sanguis can degrade arginine via the arginine deiminase (AD) system. The three enzymes of this system, AD, ornithine carbamyltransferase (OTC), and carbamate kinase (CK), catalyze the breakdown of arginine to ornithine, CO2, and two molecules of ammonia, with the production of ATP from ADP. The genes of the AD system, which are subject to complex regulation in the oral streptococci, have been isolated in bacteriophage lambda by screening for AD activity. The AD gene, designated arcA, was expressed from recombinant bacteriophage or in cells harboring plasmid subclones from this phage at a level up to 1,000-fold lower than the level in fully derepressed S. sanguis but apparently under the control of its own promoter. By subcloning in Escherichia coli mutants defective in anabolic OTC (argF argL) and CK (carB), it was demonstrated that the genes for S. sanguis OTC and CK were located adjacent to the AD gene. The levels of expression of the OTC and CK genes (arcB and arcC, respectively) were also very low in E. coli, although arcC expression was not as poor as arcA and arcB expression when compared with the levels found in S. sanguis. Also, arcB and arcC were unable to complement the defects in their anabolic counterparts. Introduction of the entire AD system or subclones which encoded only the AD gene into E. coli harboring defects in arginine and pyrimidine biosynthesis resulted in a 10- to 15-fold decrease in the level of AD activity, suggesting that arginine or its metabolites may regulate AD expression. Transposon mutagenesis was utilized to construct defined mutants of S. sanguis with mutations in the AD gene cluster. AD gene expression in these mutants indicated that the expression of the AD genes in this organism is strongly interrelated. The isolation and partial characterization of the arc genes represents the first step in the genetic manipulation of the AD system in the oral streptococci for analysis of the regulation of AD, analysis of the role of the system in plaque ecology, and utilization of the system to modulate the cariogenicity of dental plaque. [TOP OF PAGE]

  16. Evolution of sex in RNA viruses. Chao, L. (1989). J. Theor. Biol. 133:99-112. The distribution of deleterious mutations in a population of organisms is determined by the opposing effects of two forces, mutation pressure and selection. If mutation rates are high, the resulting mutation-selection balance can generate a substantial mutational load in the population. Sex can be advantageous to organisms experiencing high mutation rates because it can either buffer the mutation-selection balance from genetic drift, thus preventing any increases in the mutational load (Muller, 1964: Mut. Res. 1, 2), or decrease the mutational load by increasing the efficiency of selection (Crow, 1970: Biomathematics 1, 128). Muller's hypothesis assumes that deleterious mutations act independently, whereas Crow's hypothesis assumes that deleterious mutations interact synergistically, i.e., the acquisition of a deleterious mutation is proportionately more harmful to a genome with many mutations than it is to a genome with a few mutations. RNA viruses provide a test for these two hypotheses because they have extremely high mutation rates and appear to have evolved specific adaptations to reproduce sexually. Population genetic models for RNA viruses show that Muller's and Crow's hypotheses are also possible explanations for why sex is advantageous to these viruses. A re-analysis of published data on RNA viruses that are cultured by undiluted passage suggests that deleterious mutations in such viruses interact synergistically and that sex evolved there as a mechanism to reduce the mutational load. [TOP OF PAGE]

  17. Mutual exclusion occurs in a Chlorella-like green alga inoculated with two viruses. Chase, T.E., Nelson, J.A., Burbank, D.E., Van Etten, J.L. (1989). J. Gen. Virol. 70:1829-1836. [TOP OF PAGE]

  18. Bacteriophages and their therapeutic-prophylactic use. Chernomordick, A.B. (1989). Med. Sestra. 6:44-47. [TOP OF PAGE]

  19. Identification and characterization of a plasmid encoding phage abortive infection from Lactococcus lactis ssp. lactis UC811. Coffey, A., Fitzgerald, G.F., Daly, C. (1989). Netherlands Milk and Dairy Journal 43:229-244. Identification and characterization of a plasmid encoding abortive infection from Lactococcus lactis ssp. lactis UC811.Bacteriophage infection of starter cultures used in milk fermentations is a major problem in commercial practise. A highlight of recent genetic studies has been the demonstration of a linkage between phage insensitivity mechanisms and plasmid DNA. The conjugative plasmid pCI1829 encodes abortive infection in Lactococcus lactis ssp. lactis. This was manifested by complete insensitivity (no plaques) to small isometric-headed phage and a large reduction in plaque size for prolate-headed phage. The introduction of a derivative of a similar plasmid, pCI1750, into a strain containing pCI1829 resulted in complete insensitivity against a prolate-headed phage showing that these plasmids have an additive effect against phage. The abortive infection system described here was not inhibited by temperatures as high as 40.degree. C. [TOP OF PAGE]

  20. [A mutant of Streptococcus lactis with resistance to bacteriophages isolated in Argentina and the United States of America]. de, F.S., Parada, J.L., Ledford, R.A., Solari, A., Brown, J. (1989). Revista Argentina de Microbiologia 21:1-7. Streptococcus lactis and Streptococcus cremoris have an important role in the fermentation of milk during the manufacturing of lactic products. Bacteriophages are spread in the plant environment and they may infect the starters causing technical and economic problems to the dairy industry. It is now known that the usual methods of control are not completely safe against the proliferation of phages. It is necessary therefore to include resistant strains to the phages which infect the plant. This study introduces a simple and direct method to obtain spontaneous mutants which have performed clear resistance to a certain phage. The prolate (ARG) phages used in this work St11, St13 and St15 were isolated in Argentina, on S. lactis C2 while the prolate and isometric (AM) phages phi C2, D59-1, I16-1, and G72-1, F4-1, I37-1 respectively, were isolated in Cornell University. on the corresponding S. lactis strains except the phage D59-1 isolated on an S. cremoris strain. The propagation of phages was performed using M and M17 broth. The phage sensitivity was tested through spot test and overlay plaque plating method expressing the titre of phage suspension. as PFU/ml. In order to obtain spontaneous resistant mutants of S. lactis C2 to phage St15, and active culture of S. lactis C2 (sensitive) was spread on agar M plates, inoculated with drops of a suspension of phage St15 (10(8)PFU/ml) and incubated at 30 degrees C for 48 h. Colonies of mutants grown in the lytic areas caused by the phage were isolated and selected. [TOP OF PAGE]

  21. The enumeration of F-specific bacteriophages from environmental waters. Debartolomeis, J. (1989). Rhode Island Univ., Kingston (USA). The purpose of this research is to design an Escherichia coli (F super(+)) strain for the selective enumeration of F-specific bacteriophages and evaluate their levels in comparison to other bacterial indicators in various fecally polluted waters. In most cases, phage levels in excess of 10 super(3) per 100 ml were found in the final postchlorinated effluents. Chlorination treatment, as practiced by most treatment facilities, reduced the coliform and enterococci levels nearly four orders of magnitude while the F-specific phages and Clostridium perfringens) spores were reduced by less than one log. [TOP OF PAGE]

  22. Receptor specificity of the Escherichia coli T-even phage Ox2. Mutational alterations in host range mutants. Drexler, K., Riede, I., Montag, D., Eschbach, M.L., Henning, U. (1989). J. Mol. Biol. 207:797-803. [TOP OF PAGE]

  23. Plaque assay for virulent Legionella pneumophila. Fernandez, R.C., Lee, S.H., Haldane, D., Sumarah, R., Rozee, K.R. (1989). Journal of Clinical Microbiology 27:1961-1964. Methods of assessing virulence of Legionella pneumophila, the etiologic agent of Legionnaires disease, include the infection of guinea pigs, fertile chicken eggs, and mammalian and protozoan cell cultures. Guinea pig assays, in particular, are expensive, laborious, or unsuitable for routine screening of Legionella isolates. We have developed a virulence assay that requires the enumeration of viruslike plaques which are the result of virulent L. pneumophila infecting mouse L929 cells. Each plaque is the consequence of the initial infection of an L cell with a single bacterium. A nonvirulent mutant derived from the serial passage of virulent L. pneumophila on Mueller-Hinton agar fails to survive within L cells and consequently fails to produce plaques. [TOP OF PAGE]

  24. Phage ecology [book review]. Ford, R.E. (1989). BioScience 39:189-190. [TOP OF PAGE]

  25. The effect of IncP-2 plasmids on propagation of Pseudomonas aeruginosa bacteriophages. Freizon, E.V., Kopylova, Yu.I., Cheremukhina, L.V., Krylov, V.N. (1989). Genetika 25:1168-1178. [TOP OF PAGE]

  26. Molecular characterization and expression in Escherichia coli of the gene complex encoding the polysaccharide capsule of Neisseria meningitidis group B. Frosch, M., Weisgerber, C., Meyer, T.F. (1989). Proc. Natl. Acad. Sci. USA 86:1669-1673. The gene complex encoding all determinants of the biosynthesis pathway of the capsule of group B meningococci (cps) has been cloned in Escherichia coli. A 24-kilobase large chromosomal fragment is necessary for capsule expression on the E. coli surface. By transposon and deletion mutagenesis, two separate steps in transport of the polysaccharide from the cytoplasm to the periplasm and further to the cell surface became evident. Mutants were also isolated that accumulate soluble poly(sialic acid) in the cytoplasm. The cloned cps complex conferred to E. coli strain GC6 sensitivity for E. coli K1-specific phages; phage sensitivity was enhanced in two distinct classes of cps mutants. Southern blot experiments revealed homology to some or all other Neisseria meningitidis capsular types and other Neisseria species, depending on the fragment of the cps complex used as probe. [TOP OF PAGE]

  27. Application of gene probes to virus detection in water. Gerba, C.P., Margolin, A.B., Hewlett, M.J. (1989). Water Science & Technology 21:147-154. [TOP OF PAGE]

  28. Unbalanced growth as a normal feature of development of Bdellovibrio bacteriovorus. Gray, K.M., Ruby, E.G. (1989). Archives of Microbiology 152:420-424. In this study we have investigated the rates and spatial patterns of chromosome replication and cell elongation during the growth phase of wild-type facultatively prey-independent mutant strains of Bdellovibrio bacteriovorus. For the facultatively prey-independent mutants, the total DNA content of synchronously growing cultures was found to increase exponentially, as the multiple chromosomes within each filamentous cell replicated simultaneously. Cell mass, measured as total cellular protein, also increased exponentially during this period, apparently by means of multiple elongation sites along the filament wall. The relative rates of DNA and protein synthesis were unbalanced during growth, however, with the cellular concentration of DNA increasing slightly faster than that of protein. The original cellular DNA: protein ratio was restored in the progeny cells by continued protein synthesis during the septation period that follows the termination of DNA replication. Because of technical problems, these experiments could not be conducted on the wild-type cells, but similar results are assumed. This unusual pattern of unbalanced growth may represent an adaptation by bdellovibrios to maximize their progeny yield from the determinate amount of substrate available within a given prey cell. [TOP OF PAGE]

  29. ISOLATION AND CHARACTERIZATION OF TWO ENDOGENOUS PHAGES OF RHODOBACTER-SPHAEROIDES. HEITEFUSS, S., GIFFHORN, F. (1989). Journal of General Microbiology 135:911-920. Isolation and characterization of two endogenous phages of Rhodobacter sphaeroides.Two new temperate bacteriophages for Rhodobacter sphaeroides, designated .vphi.RsA and .vphi.RsD, were isolated from a stock of phage .vphi.RsG1 using R. sphaeroides strain DSM 159-2 as the indicator strain. Electron-microscopic examination showed that phage .vphi.RsA belonged to group B and phage .vphi.RsD belonged to group A of Bradley's morphological classification. Phage .vphi.RsA had a polyhedral head (50 nm in diameter) and a long, flexible, non-contractile tail (250 by 11 nm). Phage .vphi.RsD also had a polyhedral head (56 nm in diameter) and a sheathed contractile tail (160 by 18 nm). Both phages contained double-stranded DNA with cohesive ends. The size of the .vphi.RsA genome was about 40 kb and the G + C content was 72.4 mol%. The size of the .vphi.RsD genome was about 56 kb and the g + C content was approximately 72.6 mol%. Biotinylated DNA probes of phage .vphi.RsA and of phage .vphi.RsD hybridized to genomic DNA from R. sphaeroides strains Y and Si 4 but not to DNA from other sources. The host ranges of the two phages were determined using 26 R. sphaeroides strains and six strains of Rhodobacter capsulatus. Phage .vphi.RsA formed plaques on R. sphaeroides DSM 159-2 only, while phage .vphi.RsD plaqued on another six strains of R. sphaeroides but not on R. capsulatus. [TOP OF PAGE]

  30. A note on the isolation and propagation of lytic phages from Streptococcus uberis and their potential for strain typing. Hill, A.W., Brady, C.A. (1989). J. Appl. Bacteriol. 67:425-432. Thirty-eight of 98 strains of Streptococcus uberis were shown to be carrying lysogenic phage. Although propagating strains were rare, host modification by field strains sensitive to phage was used to increase the lytic spectra. When 120 nationally-collected strains were challenged with 25 phages, selected on the basis of differing lytic spectra and propagating strains, 30% were susceptible to at least one phage, increasing to 42% when 480 strains from a single farm were considered. A typing system based on susceptibility to lytic phage was considered feasible. [TOP OF PAGE]

  31. Localization, cloning, and expression of genetic determinants for bacteriophage resistance (Hsp) from the conjugative plasmid pTR2030. Hill, C., ROMERO, D.A., McKenney, D.S., Finer, K.R., Klaenhammer, T.R. (1989). Appl. Environ. Microbiol. 55:1684-1689. Genetic determinants for a bacteriophage resistance mechanism (Hsp+) encoded by plasmid pTR2030 (46.2 kilobases [kb]) were localized by mapping an 11.5-kb deletion that accompanied the transition of Lactococcus lactis LMA12-4 transconjugants (M. E. Sanders, P. J. Leonard, W. D. Sing, and T. R. Klaenhammer, Appl. Environ. Microbiol. 52:1001-1007, 1986) from phage resistance to phage sensitivity. The deleted 34.7-kb replicon (pTR2023, Hsp-) retained its conjugative ability, demonstrating that the phage resistance and conjugal transfer determinants were genetically distinct. The Hsp region of pTT2030, which was contained within a 13.6-kb BglII fragment, was cloned into the BamHI site of bacteriophage lambda EMBL3, and Hsp was subcloned into the Escherichia coli-Streptococcus shuttle vector pSA3. The recombinant plasmids pTK6 and pTK9 were recovered in E. coli HB101 and contained a 13.6-kb insert in opposite orientations. L. Lactis MG1363 transformants carrying pTK6 or pTK9 exhibited a significant reduction in plaque size, in addition to a slight reduction in the efficiency of plaquing for both prolate and small isometric phages. Phenotypic reactions observed for the recombinant plasmids suggest that pTR2030-encoded Hsp acts similarly against both prolate and small isometric phages. Tn5 mutagenesis was used to define the region essential for the expression of the Hsp+ phenotype. Any of four insertions within a 3-kb region resulted in the loss of phage resistance, whereas a further 26 insertions outside this locus had no effect on Hsp expression. In vitro deletion analysis confirmed that the 3-kb region contained all the information necessary for the observed resistance. [TOP OF PAGE]

  32. ??? Hurst, C.J., Benton, W.H., McClellan, K.A. (1989). Can. J. Microbiol. 35:474-??? [TOP OF PAGE]

  33. Selection and evolution of bacteriophages in cellstat. Husimi, Y. (1989). Adv. Biophys. 25:1-43. Objectives of this work were as follows: 1. to establish a laboratory experimental system utilizable in a biophysical approach to molecular evolution; and 2. to provide real world parameters to theories of molecular evolution, especially to Eigen's theory of quasi-species. Secretion type bacteriophage fd of E. coli, closely related phages and artifactual chimera phages of fd, and a virulent Q beta of E. coli were cultured continuously in a specially designed fermenter called a "cellstat". A phage is cultured in a flow of host bacterial cells. Due to its high dilution rate, the mutant cell could not be selected in a cell stat. It was therefore recognized that the cellstat is suitable for study of the selection and evolution process of a bacteriophage . . . [ABSTRACT NOT COMPLETE]. [TOP OF PAGE]

  34. Bacteriophage prevention and control of harmful plant bacteria US PATENT-4828999. MAY 9 1989. Jackson, L.E. (1989). Official Gazette of the United States Patent and Trademark Office Patents 1102:1190 [TOP OF PAGE]

  35. Bacteriophages of lactic acid bacteria. Jarvis, A.W. (1989). J. Dairy Sci. 72:3406-??? [TOP OF PAGE]

  36. RESISTANCE AGAINST INDUSTRIAL BACTERIOPHAGES CONFERRED ON LACTOCOCCI BY PLASMID PAJ1106 AND RELATED PLASMIDS. Jarvis, A.W., Heap, H.A., Limsowtin, G.K.Y. (1989). Appl. Environ. Microbiol. 55:1537-1543. Resistance against industrial bacteriophages conferred on lactococci by plasmid pAJ1106 and related plasmids.Plasmid pAJ1106 and its deletion derivative, plasmid pAJ2074, conferred lactose-fermenting ability (Lac) and bacteriophage resistance (Hsp) at 30.degree.C to Lac- proteinase (Prt)-negative Lactococcus lactis subsp. lactis and L. lactis subsp. lactis var. diacetylactis recipient strains. An additional plasmid, pAJ331, isolated from the original source strain of pAJ1106, retained Hsp and conjugative ability without Lac. pAJ331 was conjugally transferred to two L. lactis supsp. lactis and one L. lactis subsp. cremoris starter strains. The transconjugants from such crosses acquired resistance to the phages which propagated on th e parent recipient strains. Of 10 transconjugant strains carrying pAJ1106 or one of the related plasmids, 8 remained insensitive to phages through five activity test cycles in which cultures were exposed to a large number of industrial phages at incubation temperatures used in lactic casein manufacture. Three of ten strains remained phage insensitive through five cycles of a cheesemaking activity test in which cultures were exposed to approximately 80 different phages through cheesemaking temperatures. Three phages which propagated on transconjugant strains during cheesemaking activity tests were studied in detail. Two were similar (prolate) in morphology and by DNA homology to phages which were shown to be sensitive to the plasmid-encoded phage resistance mechanism. The third phage was a long-tailed, small isometric phage of a type rarely found in New Zealand cheese wheys. The phage resistance mechanism was partially inactivated in most strains at 37.degree.C. [TOP OF PAGE]

  37. Inhibitory effect of the extracts of Zingiber species on the adsorption and replication of phage LPP-1 in cyanobacterium. Jido, E.P., Dhaliwal, A.S. (1989). Toronto, Ont. (Canada). Annu. Meet. of the Phycological Soc. of America. 1900.Rhizome extracts from Zingiber officinale and Zingiber zerumbet were prepared by grinding in saline. Cyanobacterium was treated with each of the extracts (20% v/v). Extract treated cyanobacterium were inoculated with phage LPP-1. The extract of Z. officinale caused 59% inhibition of virus adsorption and 77% inhibition of burst size, whereas Z. zerumbet caused 67% inhibition of virus adsorption and 98% inhibition of burst size. [TOP OF PAGE]

  38. Occurence of bacteriophages infectecting Bacteriodes fragilis and other viruses in polluted marine sediments. Jofre, J., Blasi, M., Bosch, A., Lucena, F. (1989). Wat. Sci. Tech. 21:15-19. Numbers of B. fragilis bacteriophages in comparison to coliphages, enteroviruses and rotaviruses were evaluated by different methods in sediments of a coastal area near Barcelona which received substantial amounts of pollution of domestic origin. Phages infecting B. fragilis should be eluted from sediments prior to their enumeration, in the same way as solid-associated animal viruses. Phages infecting B. fragilis were better eluted by glycine buffer at alkaline pH than by a caotropic agent (beef extract-sodium nitrate). Such differences between glycine buffer and sodium nitrate were not evident when enteroviruses and rotaviruses were eluted from sediments. This suggests that elution with glycine buffer is preferable for bacteriophages, while teh use of caotropic is advisable for animal viruses, because of the simplicity of the methodology. In the studied arte, coliphages were the more abundant viruses. B. fragilis phages outnumbered rotaviruses and enteroviruses bya factor of more than ten. The ratios between phages active against B. fragilis and either enteroviruses or rotaviruses in marine sediments were similar to the ratios found in sewage, thus indicating that they have a similar fate. [TOP OF PAGE]

  39. Determinants of the efficacy of tobramycin therapy against isogenic nonmucoid and mucoid derivatives of Pseudomonas aeruginosa PAO1 growing in peritoneal chambers in mice. Kelly, N.M., Rawling, E.G., Hancock, R.E. (1989). Antimicrobial Agents & Chemotherapy 33:1207-1211. Mice which were supporting the growth of Pseudomonas aeruginosa in chambers implanted in their peritoneums were given two intramuscular injections of tobramycin (15 mg/kg of body weight each) at an interval of 8 h. Three hours later, chambers were removed and their contents were assessed for viable counts. Controls revealed that tobramycin levels in the chambers were 3.8 micrograms/ml 15 min after injection of 15 mg of tobramycin per kg and remained above the in vitro MICs (0.5 to 1 microgram/ml) for the tested strains for 8 h. It was demonstrated that tobramycin therapy was less effective with higher inocula and with longer delay before administration. Thus, in vivo, the concentration of bacteria in the chambers at the time of the first tobramycin injection had a profound effect on the bactericidal efficacy of tobramycin therapy. No such concentration dependence was observed in mock in vitro therapy experiments. A phage-selected mucoid derivative of P. aeruginosa PAO1 showed only a marginal increase in in vitro aminoglycoside susceptibility and no major alteration in in vivo susceptibility compared with its isogenic nonmucoid parent strain. [TOP OF PAGE]

  40. Inducible bacteriophages from ruminal bacteria. Klieve, A.V., Hudman, J.F., Bauchop, T. (1989). Appl. Environ. Microbiol. 55:1630-1634. [TOP OF PAGE]

  41. [Phagotherapy of postoperative suppurative-inflammatory complications in patients with neoplasms]. [Russian]. Kochetkova, V.A., Mamontov, A.S., Moskovtseva, R.L., Erastova, E.I., Trofimov, E.I., Popov, M.I., Dzhubalieva, S.K. (1989). Sovetskaia Meditsina 23-26. The authors assess the efficacy of phage therapy ofsuppurative and inflammatory complications in oncological patients. A clinical and laboratory analysis has involved 131 patients whose etiotropic therapy consisted of bacteriophages (65 patients) and antibiotics (66). Medicinal phages, manufactured by the Tbilisi Research Institute for Vaccines and Sera, have been administered according to 3 schemes: (1) parallel with antibiotics, (2) after long ineffective antibiotic therapy, (3) phages alone starting from the onset of the purulent complication. The preparations have been prescribed with due consideration for the isolated microflora sensitivity. Incorporation of phages in combined therapy of infectious complications has yielded positive results in 81.5% of cases, whereas antibiotics have proved effective in but 60.6%. The efficacy of phage therapy depends on the type of pyoinflammatory complications (the results are the best in the management of wound infections), the microflora pattern of the purulent foci (phages are the most effective with a corresponding monoinfection), characteristics of the therapeutic phages proper (Pseudomonas aeruginosa phage is characterized by the highest therapeutic activity, as compared to staphylococcal and other phages). [TOP OF PAGE]

  42. Gene Transfer in the Environment. Levy, S.B., Miller, R.V. (1989). McGraw-Hill, New York.[TOP OF PAGE]

  43. ENHANCEMENT OF BACTERIOPHAGE PHI-X-174 PLAQUES BY HOMOIONIC CLAY MINERALS. LIPSON, S.M., ALSMADI, O.A. (1989). Journal of General Microbiology 135:3497-3504. Enhancement of bacteriophage .phi.X-174 plaques by homoionic clay minerals.The incorporation of selected particulates or charged polymers into qualititative and quantitative assay systems has been found to increase virus specific infectivity. The purpose of this study was to identify a mechanism(s) to explain the effect of charged particulates upon plaquing efficiency. The clay minerals kaolinite (K), montmorillonite (M), with the bacteriophage .vphi.X-174 and its host Escherichia coli, were used as a model system. Enhanced bacteriophage infectivity (expressed as plaque-forming units) occurred at K and M concentrations in the agar overlay of 0.14-0.50 and 0.01-0.15 mg ml-1, respectively. A maximum increase of bacteriophage titre (to 180-230% of the control value) occurred at 0.18-0.20 and 0.06-0.08 mg ml-1 K and M, respectively. Increased infectivity was related to the type of clay mineral and to the cation saturating the exchange complex in the order K homoionic (i.e. saturated with a single cation type) to Na+ > Mg2+> K+ and M homoionic to Mg2+ > Na+ > K+. Enhanced infectivity of the bacteriophage was not related to the surface area of K nor to the external or total surface area of M. The increase of bacteriophage titer above that of the control was only minimally related to cation-exchange capacity, but was asssociated with the anion-exchange capacity of the clay. The increased plaquing efficiency is ascribed to the formation of high-density bacteria-clay microcosms which, in turn, modify bacteriophage contact and adsorption to the host cell. [TOP OF PAGE]

  44. Use of a bacteriophage-encoded glycanase enzyme in the generation of lipopolysaccharide O side chain deficient mutants of Escherichia coli O9:K30 and Klebsiella O1:K20: role of O and K antigens in resistance to complement-mediated serum killing. McCallum, K.L., Laakso, D.H., Whitfield, C. (1989). Canadian Journal of Microbiology 35:994-999. Coliphage K30 lysates contain free and phage-associated forms of a bacteriophage-encoded capsule depolymerase (glycanase) enzyme, active against the serotype K30 capsular polysaccharide of Escherichia coli. The free glycanase has been purified to apparent homogeneity. The molecular weight of the enzyme was estimated at 450,000, and when heated in SDS at 100 degrees C, the enzyme dissociated into two subunits of 90,000 and 52,000. The glycanase enzyme was used as a reagent to reversibly degrade the capsular layers on cells of Escherichia coli O9:K30 and Klebsiella O1:K20. This treatment rendered these bacteria sensitive to their respective lipopolysaccharide-specific bacteriophages, coliphage O9-1 and Klebsiella phage O1-3. This novel approach facilitated isolation of lipopolysaccharide O antigen side chain deficient mutants which retained the ability to synthesize the capsule. The response of defined mutants, O+:K-, O-:K+, and O-:K-, to exposure to nonimmune rabbit serum was measured. Results showed that the primary barrier against complement-mediated serum killing in both Escherichia coli O9:K30 and Klebsiella O1:K20 was the O antigen side chains of the lipopolysaccharide molecules. In both strains, the capsule played no role in the determination of serum resistance. [TOP OF PAGE]

  45. Functional relationships and structural determinants of two bacteriophage T4 lysozymes: A soluble gene e and a baseplate-associated gene 5 protein. Mosig, G., Lin, G.W., Franklin, J., Fan, W.H. (1989). New Biologist 1:171-180. Lysozymes have proved useful for analyzing the relation between protein structure and function and evolution. In bacteriophage T4, the major soluble lysozyme is the product of the e gene, gpe (gene product = gp). This lysozyme destroys the wall of its host, Escherichia coli, at the end of infection to release progeny particles. Phage T4 contains two additional lysozymes that facilitate penetration of the baseplates into host cell walls during adsorption. At least one of these, a 44-kD protein, is encoded by gene 5. We show here that a segment of the gp5 lysozyme amino acid sequence, deduced from the DNA sequence of gene 5, is remarkably similar to that of the T4 gene e lysozyme. Both T4 lysozymes are somewhat similar to the lysozyme of the Salmonella phage P22, but there is little significant DNA sequence homology among the two T4 lysozyme genes and the P22 lysozyme gene. We speculate that these lysozymes are adapted to differences in the composition of the cell walls of E. coli and S. typhimurium. The cloned gene 5 of phage T4 directs synthesis of a 63-kD precursor protein that is approximately 19 kD larger than the gene 5 protein isolated from baseplates. Gp5 first associates with gp26 to form the central hub of this structure. Since gp26 synthesized from the cloned gene is approximately 19 kD smaller than the 26 protein found in the hub, we speculate that a peptide cleaved from gp5 is transferred to gp26 during assembly and that this processing is important in assembly and function of this complex protein machine and in modulation of the activity of the 44-kD baseplate lysozyme. [TOP OF PAGE]

  46. Coliphages as indicators of faecal pollution at three recreational beaches on the Firth of Forth. O'Keefe, B., Green, J. (1989). Water Res. 23:2696-2701. [TOP OF PAGE]

  47. [Research, education and environmental health related to pollution in the Gulf]. Paoletti, A., Parrella, A., Gargiulo, E., Aliberti, F. (1989). Annali di Igiene 1:495-523. At Italian-Russian International Conference on "Role of the University on ecological education and training," was illustrate six topics of 30 years of our scientific and didactic activity on Environmental Hygiene, here below summarized: I. At first time, sludges of biological treatment plants and domestic sewage were frequently utilized under bacteriological control as economical and ecological fertilizers of land and waters. At present such a custom is very rare owing the chemical pollution of sewage continuously increasing; but in some countries it is still in use, and is our opinion and experience that organic waste material must be reused as fertilizer of land, more and more devoid of humus and subject to erosion of winds and waters. Some treatment plants are shown, and related plankton pyramid's. II. Pilot sewage treatment plants are frequently used in our experiments and training, for study and control of the biological degradation of organic matter, for evaluate the disappearance rate of bacteria and viruses, for investigate the foric action and behaviour of chemical and radioactive pollutants, for quantify their accumulation in the sludges of sewage treatment plants, and so forth. Different pilot plants are used, located both in our laboratories both in industries; in nuclear power plant are tested at the same time 3 models of 3 different plants (biodiscs, activated sludges, biofiltering channels), working with prevalent algal growth. III. Many species of microorganisms (metazoa, protozoa, algae, fungi, bacteria, viruses and new species like Bdellovibrio) are present in aerobic sewage treatment plants (activated sludge, bacterial bed, biodiscs, lagooning, etc.); in anaerobic treatment (digesters) prevail only methane-producing bacteria. Some of these organisms are very abundant and very active as consumers of organic matter; others are characteristic indicators of well balanced purification or of bad purification owing acute variation of organic load or presence of toxic substances in sewage. Many strains are antibiotic-producing, or Vit. B12 producing; others explain a strong lytic activity or are neuraminidase-formers. Production of great amount of biofloculant polysaccharides useful on sedimentation of organic matter is enhanced by adding particular organic pollutants like distillery wastes and others. Sewage treatment plants are good means for scientific research of particular biota continuously available and for food microbiological training for students and technicians on pathogens present in treated and untreated sewage and in sludges. IV. Big fecal pollution of coastal waters is clearly dangerous because of bathing beaches, shellfish farming, bacterial aerosols, damage to marine biota, eutrofication, aesthetic problems. [TOP OF PAGE]

  48. In situ morphologies of some bacteria from New Zealand hot springs. Patel, B.K.C., Chalcroft, J.P., Morgan, H.W., Daniel, R.M. (1989). Systematic and Applied Microbiology [SYST. APPL. MICROBIOL. ] 11:187-193. Conventional and unconventional microbial morphotypes were observed in geothermal springs when in situ electron microscopy methods were used. The conventional morphologies observed were rod, coccal and spiral shaped cells. In certain cases, phage and flagellated cells were also observed. The unconventional morphotypes consisted of cells with "flap" like structures or filamentous cells which possessed swollen sac-like structures. Based on the typical cell wall architecture some of the microorganisms could tentatively be assigned to archaebacterial species or Thermus species. The methods described in the paper have aided in the identification and thereby the isolation of some of the unique morphotypes whereas some other types still elude isolation. [TOP OF PAGE]

  49. CONCENTRATION OF GIARDIA-LAMBLIA CYSTS LEGIONELLA-PNEUMOPHILA CLOSTRIDIUM-PERFRINGENS HUMAN ENTERIC VIRUSES AND COLIPHAGES FROM LARGE VOLUMES OF DRINKING WATER USING A SINGLE FILTRATION. Payment, P., BERUBE, A., PERREAULT, D., Armon, R., TRUDEL, M. (1989). Canadian Journal of Microbiology 35:932-935. 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-.mu.m 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. [TOP OF PAGE]

  50. Studies on Lytic Bacteriophages Lactic Streptococci. Powell, I.B. (1989). University of Melbourne (Australia). The group N lactic streptococci (lactococci) used in the manufacture of fermented milk products are susceptible to lytic bacteriophage infection. Two morphological types of phages are commonly isolated form infected dairy fermentations. Both types have a polyhedral head (isometric or prolate) and an apparently non-contractile tail. Eight prolate-headed and three isometric-headed phages (all Australasian isolates) were compared via electrophoretic analysis of virion proteins, DNA:DNA hybridization and estimation of genome size. Virion protein patterns of the prolate-headed phages were all almost identical; patterns of the isometric-headed phages were very similar to each other but unlike the patterns of prolate-headed phages. DNA:DNA hybridizations showed homology between DNAs of morphologically similar phages but not between DNAs of different phage types. Genome sizes (prolate-headed phages, 21.8 to 22.9 kbp; isometric-headed phages, 28.8 to 30.4 kbp) were also different for the two phage types. These results confirmed that the two phage types are structurally and genetically distinct. Restriction site maps of prolate phage DNAs showed that some of the phages are closely related. Prolate-headed phage c6A infects a range of strains including Streptococcus lactis C6. Detailed study of c6A showed that it carries double-stranded DNA (21.9 kbp; G+C content 36.7%) with 5$/sp/prime$-protruding cohesive termini. This DNA was remarkably resistant to cleavage by restriction endonucleases. Molecular cloning of 34% of the phage genome in Escherichia coli plasmids revealed a significant paucity of sites for various restriction enzymes in c6A DNA. Replication of c6A involves breakdown of host cell DNA and incorporation of breakdown products into phage DNA. Host DNA breakdown and phage DNA synthesis both being within the first 3 to 6 minutes after infection. Phage DNA synthesis continues at an approximately constant rate throughout the latent period. In tryptone-yeast extract broth at 30$/sp/circ$C cell lysis begins 25 minutes after infection. The phage burst is about 124 p.f.u. per infected cell. [TOP OF PAGE]

  51. Close relationship of virulent bacteriophages of Streptococcus salivarius spp thermophilus at both the protein and the DNA level. Prevots, F., Relano, P., Mata, M., Ritzenthaler, P. (1989). Journal of General Microbiology 135:3337-3344. Comparisons of 23 virulent phases of Streptococcus salivarus subsp. thermophilus were made based on morphology, host-range structural protein analysis, DNA restriction patterns and DNA homology. All the phages had isometric heads (diameters 55-58 nm) and striated tails (lengths 221-275 nm). The genome sizes ranged from 37 to 43 kb. The electrophoretic profiles of the structural proteins were related, with at least two major bands of about 23 and 29 kDa in common for all the phages except three, .vphi.80, .vphi.96 and .vphi.99. These latter phages, which had a particular protein composition (main proteins of 29 and 40 kDa) and high DNA homology between each other, showed only a low DNA homology with the other phages. Extensive DNA homologies were demonstrated by Southern hybridization for all the other phages. All these results suggest a close evolutionary relationship between the phages. [TOP OF PAGE]

  52. The inconsistent distribution of introns in T-even phages indicates recent genetic exchange. Quirk, S.M., Bell-Pedersen, D., Tomaschewski, J., Ruger, W., Belfort, M. (1989). Nucl. Acids Res. 17:301-315. [TOP OF PAGE]

  53. COLIPHAGES AS POSSIBLE INDICATORS OF COLIFORMS IN SEWAGE. Qureshi, A.A., Qureshi, M.A. (1989). Microbios Letters 41:77-82. Coliphages as possible indicators of coliforms in sewage.Higher concentrations of coliforms, faecal coliforms and coliphages were observed during January and February in Bahrain with average daytime temperatures of 20.degree. C. Since faecal coliforms and coliphages exhibited a linear relationship in the raw sewage, it is suggested that detection of only the coliphages may be employed as an indicator of faecal pollution. [TOP OF PAGE]

  54. Evaluation of Salmonella typhimurium WG49 host assay method for enumeration of male-specific coliphages in an estuarine environment. Rhodes, M.W., Kator, H. (1989). Williamsburg, VA (USA). #1990 Annu. Meet. of the National Shellfisheries Association. 1900.An assay method for enumeration of male-specific coliphages in sewage was evaluated in a Virginia subestuary subject to nonpoint pollution sources including fecal inputs from livestock. Phages were enumerated using the bacterial host Salmonella typhimurium WG49 modified to produce Escherichia coli sex pili. WG49 has been reported to detect male-specific ribonucleic acid (FRNA) coliphages in sewage with little interference from somatic salmonella phages. FRNA phages and fecal coliforms were enumerated from water and sediment samples collected seasonally from the estuary and feeder streams and microbiological densities related to selected environmental parameters. Examination of 300 purified phage isolates showed 99% were RNAase resistant, 97% were lytic on the female parent salmonella strain (WG45), 4% were lytic on male E. coli and none were lytic on female E. coli . Parallel enumeration of samples on WG45 and WG49 yielded equal or greater phage densities on the former host. The significance of somatic phage recovery by the S. typhimurium WG49 host in an estuary lacking a point source of sewage is discussed. [TOP OF PAGE]

  55. BACTERIOLOGICAL ASPECTS OF DRINKING WATER IN AREA OF BRINDISI ITALY. Ricci, M., Aliquo, M.R., Bottinelli, G.P. (1989). Igiene Moderna 92:1133-1150. Bacteriological aspects of drinking water in area of Brindisi [Italy].The authors carried out a study on additional bacteriological parameters provided for DPCM 41 8-2-85. They noted negative results for Escherichia coli Bacteriophage and pathogenic Enterobacteria and positive results for Staphylococcus aureus and Pseudomonas aeruginosa. Nevertheless the nature of delivered water is fairly good according to fecal indexes. [TOP OF PAGE]

  56. Ecologists wary about environmental releases. Roberts, L. (1989). Science 243:1141-1141. [TOP OF PAGE]

  57. [Heterogeneity of a Pseudomonas bacteriophage population]. [Russian]. Romashko, A.M., Bylinskii, A.F. (1989). Mikrobiologiia 58:71-75. Twenty-five isolates of virulent bacteriophages for Pseudomonas belonging to the morphological group C (Bradley, 1967) were obtained from different natural habitats. The phages of each isolate were found to differ from one another in at least one of the following characteristics: the sensitivity to an osmotic shock, to heating at 60 degrees C and to UV; the ability to cause lysis of 86 Pseudomonas strains; the reaction of neutralisation with antisera. At the same time, the phages were related by the existence of common antigens. These properties are responsible for the genetic stability of the bacteriophages in their complicated relationship with their host, Pseudomonas bacteria. [TOP OF PAGE]

  58. ANTIBIOTIC RESISTANCE, HEAVY METAL RESISTANCE, CHLORINE RESISTANCE AND PHAGE TYPING PATTERNS OF FECAL COLIFORMS ISOLATED FROM SECONDARY EFFLUENT (GROUNDWATER). Rusin, P.A. (1989). University of Arizona. Antibiotic resistance profiles of fecal coliform isolated from unchlorinated and chlorinated secondary effluent were determined. Of 332 fecal coliforms isolated from chlorinated effluent a mean of 48% were multiply antibiotic resistant. In contrast, of 347 fecal coliforms isolated from unchlorinated effluent a mean of 29% were multiply antibiotic resistant. Resistance to ampicillin, cephalothin, and carbenicillin were significantly higher in the former than the latter. Randomly selected isolates survived and/or grew in sterile and unsterile effluent retaining resistance patterns for 40 days. Resistance factors were transferred in laboratory medium at frequencies from 0 to 1.2 $/times$ 10$/sp[-2]$ (number of recombinants/number of recipients) and in sterile neutralized tertiary effluent at frequencies from 0 to 1.0 $/times$ 10$/sp[-4]$. Resuscitative techniques were necessary for optimal recovery of fecal coliforms from effluent using selective media. Antibiotic resistance patterns of fecal coliforms isolated from unchlorinated and chlorinated effluent was not associated with chlorine or heavy metal resistance. Multiply antibiotic resistant fecal coliforms from chlorinated effluent were significantly less sensitive to lytic phage than multiply antibiotic sensitive fecal coliforms from unchlorinated effluent (p $<$.05). Using group discriminate analysis of data, phage typing techniques were shown to be a potential tool for tracing fecal contamination of groundwater. [TOP OF PAGE]

  59. Control of microbiofouling using bacteriophage 2. Detection of phages and fundamental study of their lytic effect on fouling bacteria. Sakaguchi, I., Shinshima, K., Kawaratani, K., Sugai, O. (1989). Denryoku Chuo Kenkyusho Hokoku 1-32. Microbiofouling of condenser tubes of thermal power plants markedly reduces cooling efficiency. Recently a method for controlling microbiofouling using bacteriophages has been receiving consideration. In this paper, studies of the ecological relationship between fouling bacteria and phages, and the lytic activity of phages to fouling bacteria are reported. With thirty cultures of the isolated fouling bacteria from microbial film on glass substratum immersed in flowing seawater at Sendai Bay in north Japan, we carried out the detection and isolation of phages lysing the bacteria from seawater from October 1987 to December 1988 about once per month. The results obtained were as follows: Seventeen cultures were sensitive to phages and were found to belong to Pseudomonas (12 strains), Acinetobacter - Achromobacter (3 strains), Vibrio (1 strain) and Flavobacterium (1 strain). Eighteen strains of phages were isolated. A high frequency of phage incidence was observed with the cultures of rapid growing bacteria. But, for cultures of slow growing bacteria, phages were rarely isolated. Under different M.O.I.(multiplicity of infection), the lytic activity of phage to host bacteria was studied. At M.O.I. higher than 0.1, the phage could effectively lyse host bacteria after 5-7 hr of infection. At M.O.I. lower than 0.01, the low level of bacteria concentration was maintained with only slight increase until 48 hr after infection. Various filamentous bacteria were commonly found as predominant species in condenser tube microbiofouling communities. However we were unable to grow the filamentous bacteria in the standard culture medium used in these experiments. Development of culture methods is necessary in order to develop control methods for filamentous bacteria. [TOP OF PAGE]

  60. ENUMERATION OF PSEUDOMONAS SPECIES AND PSEUDOMONAS-AERUGINOSA BACTERIOPHAGES IN DOMESTIC SEWAGE. SALLAL, A.-K.J., ABU-ALTEEN, K.H., Jafri, A.M. (1989). Microbios 60:35-44. Enumeration of Pseudomonas species and Pseudomonas aeruginosa bacteriophages in domestic sewage.Domestic sewage in Kuwait is mainly treated by an activated sludge process. Pseudomonas species were enumerated at all steps of sewage treatment. About 98-99% reduction in the number of these bacterial species were found in the treated effluent compared with raw sewage, which indicates a rather efficient removal of Pseudomonas from sewage. Spherical tail-less phages of Pseudomonas aeruginosa were found in all sewage samples. About 25-85% of the total phages encountered with the raw sewage were retained in the treated effluents. Seasonal variations of Pseudomonas spp. and P. aeruginosa phages in two treatment stations are reported. [TOP OF PAGE]

  61. Degrees of relatedness of T-even type E. coli phages using different or the same receptors and topology of serologically cross-reacting sites. Schwarz, H., Riede, I., Sonntag, I., Henning, U. (1989). EMBO Journal 2:375-380. [TOP OF PAGE]

  62. RELATIONSHIP BETWEEN TEMPERATE BACTERIOPHAGE 0241 AND VIRULENT BACTERIOPHAGE 832-B1 OF LACTOBACILLUS-HELVETICUS. Sechaud, L., CALLEGARI, M.-L., Rousseau, M., MULLER, M.-C., ACCOLAS, J.-P. (1989). Netherlands Milk and Dairy Journal 43:261-278. Relationship between temperate bacteriophage 0241 and virulent bacteriophage 832-B1 of Lactobacillus helveticus.In this study, a temperate phage, 0241, and a virulent phage, 832-B1, of L. helveticus were compared. Both phages were morphologically indistinguishable and were also similar in their protein profiles, serological characteristics and host ranges. The phage heads were isometric (53 nm in diameter) and the phage tails, fitted with contractile sheaths, were 150 nm long. Both phages showed three strong protein bands of 52, 41.5 and 23 kDa for 0241 and of 53.5, 43 and 23 kDa for 832-B1. Immunoblots and seroneutralization confirmed the close relationships of the two phages. Differences in adsorption rates on indicator strain CNRZ 892 and on strain CNRZ 241 were evident. The vegetative growth cycles of both phages at 37.degree. C were also significantly different being approximately 2h for 832-B1 and 4h for 0241, while the burst sizes were approximately 300 and 100 PFU/infectious centre, respectively. The implications of such a relationship in the applied field were discussed. [TOP OF PAGE]

  63. Gene l of bacteriophage f-X-174 codes polyfunctional lethal peptides. Shumilov, V.Y. (1989). Doklady Akademii Nauk SSSR 308:1008-1012. The in vivo identification of a gene L product was carried out in Escherichia coli. The structures of the gene L of phage f.X174 and of the adjacent plasmid pVYB215 were described, as was the homology of the structure of L-peptides with various proteins. Apparently, the gene L belongs to the group of ecological adaptation genes. These genes are not necessary for ensuring the vital activity of the phage under optimal conditions of a laboratory culture, but they are necessary to ensure the survival of the phage under changing conditions of the external and internal environment of the host cell. [TOP OF PAGE]

  64. MECHANISMS OF PHAGE RESISTANCE ENCODED BY CONJUGATIVE PLASMIDS OF LACTOCOCCI. Sing, W.D. (1989). North Carolina State University. Phage resistance mechanisms encoded by the conjugal plasmid pTR2030 from Lactococcus lactis subsp. lactis ME2 and newly identified plasmids, pTRK11, pTRK12, pTRK30, and pTRK317 from Lactococcus lactis subsp. cremoris TDM1 were studied. The objectives of this investigation were to characterize the mechanisms of phage defense, examine physical and genetic properties of the plasmids involved, and develop genetic strategies for the construction of phage-insensitive starter cultures. The effect of pTR2030 on phage DNA injection, transfection, release of progeny phage, and cell death was evaluated for a number of lactococcal phages. Although phage DNA infection was not affected, pTR2030 reduced phage burst size and the number of infected hosts which generated viable phage. The data confirmed that pTR2030 interferes with development of prolate and small isometric phages via a classical abortive infection (Hsp) mechanism. Restriction and modification (R/M) plasmids (pTRK11, pTRK12, pTRK30, pTRK317) were isolated from an industrial starter strain, Lactococcus lactis subsp. cremoris TDM1 using conjugation and transformation. The specificity of R/M conferred by three of the plasmids was different. Efficiencies of plaquing (EOP) for phage c2 on R$/sp[+]$/M$/sp[+]$ hosts carrying these plasmids was 10$/sp[-2]$ to 10$/sp[-4]$. When R/M encoded by pTRK12 or pTRK30 was combined with abortive infection (Hsp$/sp[+]$) encoded by pTR2030 in the same strain, the resistance phenotype improved or weakened depending on the specific nature of the coexisting plasmids. In one case, R/M complemented Hsp and maintained phage-infected host survival. An effective starter rotation sequence, designated the phage defense rotation strategy (PDRS), was developed using isogenic strains harboring different phage-defense systems (Hsp$/sp[+]$ or R$/sp[+]$/M$/sp[+]$). Phage-resistant derivatives carrying pTR2030 (Hsp$/sp[+]$) or R$/sp[+]$/M$/sp[+]$ plasmids of different restriction specificities were evaluated in specific rotation sequences by starter culture activity tests in milk contaminated with phage. Initiating the sequence with a Hsp$/sp[+]$ strain reduced the level of phage contamination, enabling the effective use of R$/sp[+]$/M$/sp[+]$ strains. R$/sp[+]$/M$/sp[+]$ strains suppressed development of phage resistant to the Hsp mechanism. A model sequence for the PDRS was successful through 9 cycles when challenged with commercial phage composites during starter activity tests that mimic conditions encountered during cheesemaking. [TOP OF PAGE]

  65. Genetic definition of two functional elements in a bacteriophage T4 host-range "cassette". Snyder, M., Wood, W.B. (1989). Genetics 122:471-479. [TOP OF PAGE]

  66. UV-INACTIVATION OF MICROORGANISMS IN WATER. SOMMER, R., WEBER, G., CABAJ, A., WEKERLE, J., KECK, G., SCHAUBERGER, G. (1989). Zentralblatt fuer Hygiene und Umweltmedizin 189:214-224. UV-inactivation of microorganisms in water.UV-inactivation of Escherichia coli, Bacillus subtilis spores, Staphylococcus-Phage A 994, Poliovirus type Mahoney and Rotavirus SA 11 was tested under controlled physical conditions. B. subtilis-spores were found to be the most resistant of these microorganisms, followed by Rotavirus, Bacteriophage and Poliovirus. E. coli required the lowest irradiation dose for inactivation. Causes and meaning of these dose-survival-reactions are discussed. [TOP OF PAGE]

  67. THE OCCURRENCE OF BACTERIOPHAGES IN SPIRIT VINEGAR FERMENTATION. STAMM, W.W., KITTELMANN, M., Follmann, H., TRUEPER, H.G. (1989). Applied Microbiology and Biotechnology 30:41-46. The occurrence of bacteriophages in spirit vinegar fermentation.Bacteriophages in concentrations of approximately 108 - 109 particles/ml were demonstrated in culture lysates of industrial submerged spirit vinegar fermentations running in different European vinegar factories. Besides frequently found fragments of phages, two types of phages could be described. The frequent type I seems to belong to Bradley-group A; type II, with a remarkably larger head, may belong to group C. For stimulation in the laboratory a phage-contaminated industrial culture was kept over 102 charges (time of cycle 24- > 240 h) in a semicontinuously operated 8-1 fermentor. A close relationship between spontaneous breakdowns and phage occurrence exists. Discrimination of breakdowns caused by lack of ethanol could not be made. Attempts to establish a bioassay for the phages failed presumably because of the instability of the phages. Continuing cultivation and waiting for secondary growth finally results in stable and productive fermentation. [TOP OF PAGE]

  68. Isolation and characterization of two rumen Streptococcus bovis bacteriophages. Styriak, I., Kmet, V., Spanova, A. (1989). Microbiologica (Pavia) 12:317-322. A method for the isolation of Streptococcus bovis bacteriophages from ruminal fluid of calves is described. Thirty to 2 .times. 103 phages per ml infecting Streptococcus bovis strains 4/1 and 47/3 were isolated directly from ruminal fluid. Two bacteriophages were characterized on the basis of plaque morphology, host ranges, electron microscopic morphology and DNA restriction endonuclease digestion patterns. The F1 and F3 phages formed clear plaques of different sizes. The plaque size of the F1 phage was about 1-1.5 mm in diameter, while the plaques of the F3 phage were larger (1.5-2.5 mm in diameter). Both phages are placed in group B of Bradley's scheme and have different host ranges. The first isolation of Streptococcus bovis phage DNA is reported. Restriction analysis of their DNAs showed that phages F1 ad F3 had different numbers of cleavage sites in their genomes and that they were not identical. [TOP OF PAGE]

  69. Human origins of Bacteroides fragilis bacteriophages present in the environment. Tartera, C., Lucena, F., Jofre, J. (1989). Appl. Environ. Microbiol. 55:2696-2701. [TOP OF PAGE]

  70. PLASMID PROFILES OF LACTOCOCCUS-LACTIS-SSP-CREMORIS STRAINS AND MUTANTS IN RELATION TO PHAGE RESISTANCE. VLEGELS, P.A.P., HELMERHORST, T.H., VAN, D.E.R., V, BIESTERVELD, S., Wouters, J.T.M. (1989). Netherlands Milk and Dairy Journal 43:245-260. Plasmid profiles of Lactococcus lactis ssp. cremoris strains and mutants in relation to phage resistance.Lactococcus lactis subsp. cremoris R1 and L. lactis subsp. cremoris 10 contain a large and complex plasmid complement. Plasmid profiles of bacteriophage-resistant mutants of these strains, which failed to adsorb their homologous phage in an efficient way, were examined and compared with that of their wild type strain. In addition to derivatives with plasmid profiles corresponding to that of their wild type strain, mutants with a variety of changes in plasmid composition could also be demonstrated. Some of them showed plasmid deletions, which seemed indicative of plasmid encoded phage sensitivity. Although the rather high mutation frequencies also pointed to plasmid involvement, no correlation between phage resistance and loss of certain plasmids could be demonstrated for either strain. But nevertheless, phage-sensitive derivatives of the resistant strains R1E and R1K appeared after long term sub-cultivation in batch or continuous culture with plasmid profiles corresponding to that of L. lactis subsp. cremoris R1. This observation led to the hypothesis that the phage resistance seemed reversible and was likely to be connected to instability of the genetic material of the strain. [TOP OF PAGE]

  71. Mutants of Escherichia coli O9:K30 with altered synthesis and expression of the capsular K30 antigen. Whitfield, C., Schoenhals, G., Graham, L. (1989). Journal of General Microbiology 135 ( Pt 10):2589-2599. Escherichia coli K30 produces a thermostable group I capsular polysaccharide. Two classes of mutants were isolated with defects in the synthesis or expression of capsule. The most common mutant phenotype was acapsular (K-), with no K-antigen synthesized. A second class of mutants, termed Ki or intermediate forms, produced colonies which were indistinguishable from those of acapsular forms yet K-antigenicity was expressed. Previous studies had demonstrated that E. coli strains that produce K30 antigen synthesize a lipopolysaccharide (LPS) fraction that is recognised by monoclonal antibodies against the K30 antigen. Synthesis of this LPS fraction was not affected in Ki forms. The results of morphological examination, LPS analysis and phage sensitivity studies are consistent with the interpretation that the defect in Ki strains results from an inability to polymerize the K30 antigen. Using plasmid pULB113 (RP4::mini-Mu), mutations resulting in both K- and Ki phenotypes were localized near the his region of the chromosome. [TOP OF PAGE]

  72. Yeast virology. Wickner, R.B. (1989). FASEB J. 3:2257-2265. [TOP OF PAGE]

  73. Characterization of a RNA virus from the parasite Leishmania. Widmer, G., Comeau, A.M., Furlong, D.B., Wirth, D.F., Patterson, J.L. (1989). Proc. Natl. Acad. Sci. USA 86:5979-5982. We were interested in screening a series of isolates of the protozoan Leishmania for the presence of viruses. The experimental procedure we used was based on an enzymatic assay originally developed for viral RNA-dependent RNA polymerases. Simultaneously, total promastigote nucleic acid preparations were analyzed for the presence of viral genome and/or transcripts. Two isolates, both classified as L. braziliensis guyanensis, were found to be positive for RNA polymerase activity and to carry a large (6 kilobases) RNA species. The polymerase reaction products hybridized to the 6-kilobase RNA, believed to be the viral genome. In conjunction with electron microscopical observations these results indicate the presence of an RNA virus in these Leishmania isolates. [TOP OF PAGE]

  74. Evaluation of potassium permanganate for inactivation of bacteriophage MS-2 in water systems. Yahya, M.T., Landeen, L.K., Forshoefel, N.R., Kujawa, K., Gerba, C.P. (1989). J. Environ. Sci. Hlth. A24:979-990. Evaluation of potassium permanganate for inactivation of bacteriophage MS-2 in water systems.Potassium permanganate (KMnO4) has been as an oxidant for decades to remove and control iron and manganese in surface water supplies. This oxidant was investigated for its ability to inactivate bacteriophage MS-2 and thereby reduce the amount of chlorine required for a 99.99% reduction of virus during drinking water treatment as required by the U.S. Environmental Protection Agency's Surface Treatment Rule (U.S. EPA 1989). Experiments were conducted in potassium monophosphate buffer (pH 6.0 and pH 8.0) at 7.degree. C. At time intervals from 0 to 30 min, samples were taken and mixed immediately with a solution of sodium thiosulfate:sodium thioglycolate to neutralize residual KMnO4. At 0.5 and 5.0 mg/L KMnO4, results showed no singificant differences (P.ltoreq.0.05) in the inactivation of MS-2 between experiments done at pH 6.0 and those at pH 8.0. Ninety-nine percent of the virus was inactivated after 50,35, and 5 min of exposure time to 0.5, 1.5, and 5.0 mg/L potassium permanganate at pH 8.0, respectively. It appears that at the currently used levels of KMnO4 (up to 10 mg/L), this oxidant may supplement high levels of chlorination in the disinfection of water systems. [TOP OF PAGE]

  75. Use of a biotinylated DNA probe to detect bacteria transduced by bacteriophage P1 in soil. Zeph, L.R., Stotzky, G. (1989). Appl. Environ. Microbiol. 55:661-665. [TOP OF PAGE]

  76. Isolation and identification of phages attacking Sterptomyces spiramyceticus 298-36. Zhou, J.M., Zhu, Y.X., Chen, X.K., Liu, Y.S., Zhu, J.H., Shen, R.Q., Sheng, Z.J. (1989). Chinese Journal of Antibiotics 14:247-249. Nineteen cultures of phage attacking S. spiramyceticus were isolated from the abnormal fermentation liquid and soil samples. Phages were purified and classified into four serotype according to the serological neutralization reaction. The phage plaque of 1010 and 4031 were large and translucent, and P11 and 1012 were small and transparent. Representative phages of these form serotypes, namely P11, 1010, 1012 and 4031 were studied maphologically and restriction fragment length polymorphism of their DNA was observed. [TOP OF PAGE]

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