by Jim D. Karam

Bacteriophages are extremely diverse in the range of microbial hosts that they infect and the chemical nature, size and geometry of their genetic materials. As the most abundant organisms on Earth [1], they contribute substantially to overall diversity of the gene pool in the microbial world. It is no surprise then that they have become prominent in the ongoing discussion on microbial diversity [2-4]. The interactions of phages with their hosts are not only important for maintenance of the ecological balance, they may also constitute a major component of the network for lateral transfer of genes among microorganisms. This notion is based on studies with all types of archived phages, which of course represent only a minute fraction of the estimated number of unique phage genomes in nature. Nevertheless, the extent to which phage may contribute to microbial diversity is becoming better appreciated because of an ongoing expansion of sequence databases for all types of organisms, including phage. In the US, funding agencies have teamed together to coordinate a significant increase in support for microbial genome sequencing projects, with a great deal of emphasis being placed on speed and economy of generating, assembling, annotating, and sharing sequence data. It is anticipated that the microbial genome database will continue to expand at a fast pace in the foreseeable future.

The most commonly used approach for sequencing a microbial genome involves genomic library construction (usually from randomly sheared genomic DNA), sequencing a large number of library clones (high-throughput sequencing), computer-assisted assembly of sequence data into contiguous segments (contigs), and carrying out more sequencing and data assembly to close gaps between contigs. The high-throughput stage is expected to yield data from overlapping clones for better accuracy of sequence reads and length of assembled contigs. In principle, the approach is straightforward and has the clearly defined goal of producing an accurate single contiguous sequence of the genome. In practice, it is riddled with bottlenecks that can be different for different genomes, depending on their sizes, states of modification, content of sequences that cannot be cloned and other factors. Personnel training and quality of operations in general are also critical factors to consider in such projects. It is very common for projects to use commercial outfits or collaborations with well-funded research institutes for operations that are impractical to support locally. Usually progress is very rapid during the early stages of a project, but gets bogged down in later stages. Some genomes remain 80-90% finished for months or years, but can still be mined for useful information, provided that this information is released to the scientific community. The technology continues to improve on several fronts, and we can expect that alternate approaches, e.g., circumventing cloning and more powerful computer programming, will cut down the time and expense required to produce a finished genome sequence and allow the sequencing of several genomes concurrently by the same team.

Phage oriented projects have so far completed the sequences of ~150 genomes (GenBank). In some cases, several members of the same phage family (Siphoviridae, Myoviridae or Podoviridae; ICTV nonenclature) are included in databases. Collectively, the data suggest that despite their vast differences in genetic composition, all dsDNA phages share similar genome architecture. The typical dsDNA phage genome consist of a mosaic of gene sets that are shared with other members of the same phage "genus" and gene sets that are unique to each genome and interspersed with the genus-specific sets. That is, dsDNA phage genomes seem to evolve by gathering genes from different sources, including genes that qualify the phages for membership in their particular genera. In some instances, lateral DNA transfer (by homologous or nonhomologous recombination) is suspected to be responsible for mosaic patterns that appear inside some phage genes. Since gene evolution by mutation (vertical change) and genome evolution by lateral DNA transfer probably occur independently of each other, it is difficult to relate whole genomes belonging to the same genus to one another in chronological order. Such timelines are more meaningful when sequences of shared (homologous) genes or gene clusters (or their protein products) are compared, e.g., divergence of an essential gene/protein within a phage genus. The framework represented by genomes of the T4-like phages is an excellent example of how vertical and horizontal evolution may drive diversity in a dsDNA phage genome type. The T4 genome type is large by viral standards and carries many genes that one usually finds in cellular rather than viral genomes. Among these are genes for some enzymes of intermediary metabolism, a multi-component DNA replisome, extensive machinery for genetic recombination, and certain types of mobile DNA elements (including homing endonuclease genes) that can move themselves and flanking DNA unidirectionally [5, 6]. There is also a well-studied prototype, phage T4 [7, 8], than can be used as reference when comparing nucleotide sequences and genome organization of different T4-like phages.

In a collaborative project with Henry Krisch (CNRS, Toulouse, France), we have been sequencing the genomes of a number of T4-like Myoviridae that diverge in host range and/or other characteristics, as determined by preliminary genetic and genomic scanning. Thanks to the efforts of Hans Ackermann, a number of these phages that infect bacterial hosts other than E. coli have been archived at LaValle University (Quebec, CA) and made available for our studies. The sequences of 2 Aeromonas phages, Aeh1 (A. hydrophila) and 44RR2.8t (A. salmonicida) and 2 coliphages RB69 and RB49 are now posted on a publicly accessible web site (http://phage.bioc.tulane.edu) and are in the process of being submitted to GenBank. Although the available data probably represent only a very tiny sampling of what must exist in nature for this type of phage genome, certain predictions can already be made with regards to the kind of diversity one may encounter if a much more extensive collection of T4-like phages is analyzed. For example, whereas genome size appears to be rather fixed for some dsDNA phages, T4-like genomes can vary in length over a wide range. Currently, the observed range is ~164Kbp (for phage RB49) to ~233Kbp (for phage Aeh1). So it appears that genomes of the T4 kind can recruit variable amounts of DNA to go with a certain core that is common to all. Reversible gain and loss of genes and homologues may occur depending on composition of the gene pool where exchanges take place. Based on what we know from T4 studies, the highly recombinogenic character of this genetic system may allow it to be an effective scavenger of DNA from microbial hosts. . The Aeh1 genome carries 23 tRNA genes (19 amino-acid specificities), which is one indication of DNA acquisition from cellular sources. Matches to bacterial sequences in databases account for 2-5% of the predicted ORFs for any of the genomes sequenced so far. This is probably a vast underestimate of the contribution of bacterial DNA to T4-like genomes. More likely, much of the other nonT4-like DNA we observe for these phages has its matches in microorganisms that have yet to be discovered. The combinatorial potential of the genome framework acquired by the T4-like phages might underlie a potential for these phages to cross species barriers between bacteria. If this is happening in nature, then the T4-like population and unrelated phage populations with similar potential [4] could be dynamically affecting microbial diversity on a global scale.

It is still unclear what constitutes the "core" DNA of a T4-like genome. It could be >100 ORFs. Because morphological criteria have figured significantly in the classification of phages into families and genera, it has not been surprising to find homologues of the T4 morphogenesis genes in all the genomes examined so far in the "T4-Like Genome Project" (http://phage.bioc.tulane.edu). On the other hand, homologues of the T4 DNA replication/recombination gene clusters are consistently being observed to coexist with the morphogenesis clusters. Functional coupling between replication and morphogenesis has been documented in T4 studies, and could conceivably be required for natural selection of this type of phage genome. It remains to be seen if phages of the T4 morphotype exist in nature which utilize a different mode of replication from the T4 paradigm, or vice versa. To find out, one would have to utilize specific probes to access a much larger set of genomes than exists today in laboratory archives. It is particularly important to be able to screen environmental sources for genomes of phages that cannot be isolated through traditional plaque assays. T4-like phages that have significantly larger genomes than T4 and those that grow on bacterial hosts other than E. coli (or the enterobacteria in general) are underrepresented in laboratory collections [9]. Also, no phages of this genus have been reported whose heads/genomes are much smaller than T4. Finding more of T4's relatives in a variety of environmental niches and sequencing them would boost our understanding of the pathways leading to microbial diversity. In addition, such phages/genomes would constitute a treasure chest of genes and proteins for all types of studies in basic and applied molecular biology. I beg the BEG to undertake the search for more T4-like phages.

1. Wommack, K. E., and Colwell, R. R. (2000). Virioplankton: viruses in aquatic ecosystems. Microbiol Mol Biol Rev 64, 69-114.

2. Hendrix, R. W., Smith, M. C., Burns, R. N., Ford, M. E., and Hatfull, G. F. (1999). Evolutionary relationships among diverse bacteriophages and prophages: all the world's a phage. Proc Natl Acad Sci U S A 96, 2192-2197.

3. Brussow, H., and Hendrix, R. W. (2002). Phage genomics: small is beautiful. Cell 108, 13-16.

4. Pedulla, P. L., Ford, M.E., Houtz, J.M., Karthikeyan, T., Wadsworth, C., Lewis, J.A., Jacobs-Sera, D., Falbo, J., Gross, J., Pannunzio, N.R., Brucker, W., Kumar, V., Kandasamy, J., Keenan, L., Bardarov, S., Kriakov, J., Lawrence, J.G., Jacobs jr., W.R., Hendrix, R.W., Hatfull, G.F. (2003) Origins of highly mosaic Mycobacteriophage genomes. Cell 113: 171-182.

5. Belle, A., Landthaler, M., and Shub, D. A. (2002). Intronless homing: site-specific endonuclease SegF of bacteriophage T4 mediates localized marker exclusion analogous to homing endonucleases of group I introns. Genes Dev 16, 351-362.

6. Edgell, D. R. (2002). Selfish DNA: New Abode for Homing Endonucleases. Curr Biol 12, R276-278.

7. Miller, E. S., Kutter, E., Mosig, G., Arisaka, F., Kunisawa, T., and Ruger, W. (2003). Bacteriophage T4 Genome. Microbiol Mol Biol Rev 67, 86-156.

8. Karam et al Eds., Molecular Biology of Bacteriophage T4. ASM Press, 1994.

9. Ackermann, H. W., and Krisch, H. M. (1997). A catalogue of T4-type bacteriophages. Arch Virol 142, 2329-2345.

1. Anti-viral acoustically transparent earphone cover. Ullrich, K. A. (1996). Patent number 5,545,859. An earphone cover, for attachment to an operative region of an audiometric testing device includes a body formed from a substantially antiviral, acoustically-transparent material, and is constructed for covering such operative region. The body is preferably formed as a polyethylene film with a thickness of about 1-mil. Anti-viral testing shows that the earphone cover is an effective barrier throughout a 60-minute exposure time to a viral organism described as fX174 bacteriophage ATCC# 13706-B1. Acoustic transparency testing shows the earphone cover exhibits acceptable % total harmonic distortion and attenuation. A method of preventing patient cross-contamination associated with audiometric testing is also described. Both the structure and method are usable without affecting calibration of the audiometric testing device.

2. Antibacterial therapy with bacteriophage genotypically modified to delay inactivation by the host defense system. Merril, C. R., Carlton, R. M., Adhya, S. L. (2003). Patent number 5,660,812 (see also 5,688,501; 5,660,812). The present invention is directed to bacteriophage therapy, using methods that enable the bacteriophage to delay inactivation by any and all parts of the host defense system (HDS) against foreign objects that would tend to reduce the numbers of bacteriophage and/or the efficiency of those phage at killing the host bacteria in an infection. Disclosed is a method of producing bacteriophage modified for anti-HDS purposes, one method being selection by serial passaging, and the other method being genetic engineering of a bacteriophage, so that the modified bacteriophage will remain active in the body for longer periods of time than the wild-type phage.

3. Antibacterial therapy with bacteriophage genotypically modified to delay inactivation by the host defense system together with an antibiotic. Merril, C. R., Carlton, R. M., Adhya, S. L. (1998). Patent number 5,766,892. The present invention is directed to bacteriophage therapy, using methods that enable the bacteriophage to delay inactivation by any and all parts of the host defense system (HDS) against foreign objects that would tend to reduce the numbers of bacteriophage and/or the efficiency of those phage at killing the host bacteria in an infection. Disclosed is a method of producing bacteriophage modified for anti-HDS purposes, one method being selection by serial passaging, and the other method being genetic engineering of a bacteriophage, so that the modified bacteriophage will remain active in the body for longer periods of time than the wild-type phage.

4. Bacteria and bacteriophage detection using immobilized enzyme substrates. Adams, C. A., Krejcarek, G. E., Wicks, J. H. (2002). Patent number 6,436,661. Methods of detecting bacteria including the use of an immobilized enzyme substrate, and the immobilized enzyme substrate.

5. Bacterial detection by phage transduction of ice nucleation and other phenotypes. Wolber, P. K., Green, R. L. (1995). Patent number 5,447,836. Viable bacteria may be detected in biological samples by exposing bacterial cultures obtained from the samples to transducing particles having a known host range. Such transducing particles carry a heterologous gene capable of altering the phenotype of the bacteria in a readily detectable manner. For example, the transducing particles may carry an ice nucleation gene and the alteration of phenotype may be detected using an ice nucleation assay. By employing a panel of phage, unknown bacteria may be typed based on the pattern of reactivity observed. The method is particularly useful for detecting viable bacteria which may have been debilitated by exposure to sterilizing conditions, such as in food processing. The method is also useful for tracking a target bacteria in the ambient environment.

6. Bacterial phage associated lysing enzymes for treating dermatological infections. Fischetti, V., Loomis, L. (2001). Patent number 6,248,324. The present invention discloses a composition for dermatological infections by the use of a lytic enzyme in a carrier suitable for topical application to dermal tissues. The method for the treatment of dermatological infections comprises administering a composition comprising effective amount of a therapeutic agent, with the therapeutic agent comprising a lytic enzyme produced by infecting a bacteria with phage specific for that bacteria.

7. Bacteriophage-based transgenic fish for mutation detection. Winn, R. N. (2001). Patent number 6,307,121. The present invention provides transgenic fish whose somatic and germ cells contain a genomically integrated bacteriophage lambda-derived transgene construct. The transgene construct can include an excisable test nucleic acid sequence containing a heterologous mutation target nucleic acid sequence that is detectable via bioassay in a bacterial cell into which the test nucleic acid has been introduced. The frequency of mutations in the mutation target nucleic acid sequence following exposure of the transgenic fish to one or more potentially mutagenic agents can thus be evaluated.

8. Bacteriophage-mediated gene transfer systems capable of transfecting eukaryotic cells. Chada, S., Dubensky, T. W., Jr. (1998). Patent number 5,736,388. Lamboid bacteriophage capable of specifically interacting with and delivering nucleic acid molecules to eukaryotic cells are disclosed. Such bacteriophage-derived gene transfer systems target one or more specific receptors on eukaryotic cells, for instance by incorporating mutant tail fiber proteins or by incorporating known ligands for specific eukaryotic receptors into lambda phage. Also disclosed are methods for identifying and producing modified bacteriophage tail fiber polypeptides capable of specifically interacting with eukaryotic transmembrane proteins. Methods of treating diseases using such gene transfer systems are also disclosed.

9. Bacteriophage-resistant plant growth promoting rhizobacteria. Suslow, T. V. (1986). Patent number 4,584,274. Bacteriophage-resistant plant growth promoting rhizobacteria are employed for enhancing the yield of root crops, such as potatoes, sugar beets, radishes and the like, grown in soils which are infected by bacteriophage which limit the root colonization by the corresponding wild-type rhizobacteria. The strain SH5 PR3 was deposited at the ATCC for patent purposes on Jan. 20, 1983, and granted Accession No. 39270.

10. Bacteriophage-triggered cell suicide systems and fermentation methods employing the same. Klaenhammer, T. R., Conkling, M. A., O'Sullivan, D., Djordjevic, G., Walker, S. A., Taylor, C. G. (1998). Patent number 5,792,625. Described herein is a bacterial cell containing a recombinant bacteriophage defense mechanism. The defense mechanism comprises a bacteriophage promoter (e.g., a phage f31 promoter; a T7 promoter) operatively associated with a heterologous DNA encoding a product lethal to the bacterial cell. The bacterial cell is susceptible to infection by a bacteriophage, and the promoter is activated upon the infection of said bacterial cell by that bacteriophage. Bacteria useful in carrying out the invention include both gram negative and gram positive bacteria (e.g., Lactococcus lactis; Escherichia coli); the heterologous DNA may encode an enzyme that degrades nucleic acid (e.g., the products of the LlaI restriction cassette; barnase). Recombinant DNAs useful for making the foregoing cells, cultures prepared from such cells, and fermentation methods carried out with such cells are also disclosed.

11. Bacteriophage assay. Hyman, L. J., Toth, I. K. (2003). Patent number 6,555,331. There is provided an assay suitable for the typing of bacterial strains. In the assay a predetermined amount of phage is combined with a bacterial isolate of unknown strain, the mixture being located in a suitable container. The mixture of phage and bacteria is conveniently held in a liquid or semi-liquid medium facilitating interaction of the two species. The extent of bacterial growth in the presence of the phage is measured by conventional means, preferably by means of an OD reading. Desirably the phage is retained in the selected container, which is conveniently a micro-titer plate, through use of a fixant such as 5% gelatin.

12. Bacteriophage composition useful in treating food products to prevent bacterial contamination. Averback, P., Gemmell, J. (2002). Patent number 6,461,608. The present invention is directed to novel bacteriophage compositions useful in treating food products to prevent bacterial contamination.

13. Bacteriophage genotypically modified to delay inactivations by the host defense system. Merril, C. R., Carlton, R. M., Adhya, S. L. (1998). Patent number 5,811,093. The present invention is directed to bacteriophage therapy, using methods that enable the bacteriophage to delay inactivation by any and all parts of the host defense system (HDS) against foreign objects that would tend to reduce the numbers of bacteriophage and/or the efficiency of those phage at killing the host bacteria in an infection. Disclosed is a method of producing bacteriophage modified for anti-HDS purposes, one method being selection by serial passaging of a bacteriophage, and the other method being genetic engineering of a bacteriophage, so that the modified bacteriophage will remain active in the body for longer periods of time than the wild-type phage.

14. Bacteriophage of Chlamydia psittaci. Bavoil, P. M., Hsia, R. C. (1998). Patent number 5,741,697. The present invention is directed to an isolated bacteriophage designated fCPG1. The invention is further directed to an isolated DNA molecule encoding bacteriophage fCPG1, or a fragment thereof, a DNA molecule comprising DNA encoding bacteriophage fCPG1 with heterologous DNA inserted therein, or a fragment thereof, and to oligonucleotides consisting essentially of a portion of the DNA molecule encoding fCPG1. The bacteriophage fCPG1 was isolated from Chlamydia psittaci strain Guinea Pig inclusion Conjunctivitis.

15. Bacteriophage prevention and control of harmful plant bacteria US PATENT-4828999. MAY 9 1989. Jackson, L. E. (1989). Patent number 4,828,999. For preventing or controlling bacterial harm to plants, as by disease or ice nucleation, a bateriophage composition of matter containing one or more viral h mutants specific to amutant of the bacteria concerned is produced and applied to seed, soil or soil supplements, plants, or plant materials that have been exposed to or are contaiminated with or infected by bacerial disease, or to growing plants subject to ice nucleation or other bacterial harm. The invention is concerned with the composition and with the method of producing and using same.

16. Bacteriophage resistant recombinant bacteria. Hill, C. J., Klaehammer, T. R. (1996). Patent number 5,538,864. Recombinant bacteria containing phage-encoded resistance ("Per") and methods of making and using the same are disclosed. Such bacteria are made by (a) conducting a fermentation of a substrate in a medium containing a defined bacterial culture until bacteriophage are detected in the medium, the bacteriophage being specific to at least one bacteria in the medium; (b) isolating the bacteriophage; (c) digesting DNA of the bacteriophage to produce a library of DNA fragments; (d) transforming the bacteria susceptible to said bacteriophage with the library of DNA fragments to provide transformed bacteria; (e) selecting from among the transformed bacteria, a bacteriophage-resistant transformed bacteria; (f) adding bacteriophage resistant transformed bacteria to the medium; and (g) recommencing step (a). Also disclosed are bacterial cells which contain a first bacteriophage defense mechanism (Per), wherein Per comprises a bacteriophage origin of replication (ori) operatively associated with a DNA sequence incapable of producing live bacteriophage. The bacterial cell is capable of being infected by a bacteriophage, the DNA of which, once injected into the bacterial cell, competes with Per for binding to DNA polymerase.

17. Bacteriophage RM 378 of a thermophilic host organism. Hjorleifsdottir, S., Hreggvidsson, G. O., Fridjonsson, O. H., Aevarsson, A., Kristjansson, J. K. (2002). Patent number 6,492,161. A novel bacteriophage RM 378 of Rhodothermus marinus, the nucleic acids of its genome, nucleic acids comprising nucleotide sequences of open reading frames (ORFs) of its genome, and polypeptides encoded by the nucleic acids, are described.

18. Bacteriophage, a process for the isolation thereof, and a universal growth medium useful in the process thereof. Agrawal, P., Soni, V. (2002). Patent number 6,482,632. The present invention provides a isolated bacteriophage useful as a tool for studying biological, biochemical, physiological and genetic properties of actinomycetes and other organisms which comprises a novel strain of Saccharomonospora having certain specified characteristics. The invention also relates to a process for the isolation of the said bacteriophage and/or DNA phage and to a novel universal growth medium which is particularly useful in the said process. Another embodiment of the process relates to a cloning vector which comprises a plasmid or bacteriophage comprising the phage DNA of the invention.

19. Bacteriophages as recognition and identification agents. Teodorescu, M. C., Gaspar, A. M. (1989). Patent number 4,797,363. Bacteriophages are employed as agents for recognition and identification of molecules and cellular materials, using their ability to recognize their bacterial host, by coating them with antibodies or by selecting them to perform in a manner analogous to antibodies. Visibility for identification is effected by incorporating a fluorescent agent, a radioisotope, a metal, an enzyme, or other staining material. The bacteriophage are prepared so as to bind to the molecular or cellular material through either the tail or head segment of the bacteriophage.

20. Bacteriophages, method for screening same and bactericidal compositions using same, and detection kits using same. Takahashi, S. (2001). Patent number 6,322,783. The bacteriophage has a high level of specificity to a certain specific pathogenic bacterium so that the bacteriophage can surely kill the pathogenic bacterium as a host through phogocytic [sic?] action. The bio-bactericidal material containing the bacteriophage can be applied to food such as fresh food, etc., and to places, etc. or to even persons for cooking food material such as restaurants, school kitchens, etc., or any other thing which requires disinfection from pathogenic bacteria, and it can kill pathogenic bacteria. The bio-bactericidal material containing a cocktail of two or more different kinds of the bacteriophages can kill corresponding kinds of pathogenic bacteria concurrently. Further, the phage can infect only the pathogenic bacterium as a host bacterium, and does not infect persons, making it very safe and useful.

21. Bioluminescent biosensor device. Sayler, G. S., Ripp, S. A., Applegate, B. (2003). Patent number 6,544,729. Disclosed are methods and devices for detection of bacteria based on recognition and infection of one or more selected strains of bacteria with bacteriophage genetically modified to cause production of an inducer molecule in the bacterium following phage infection. The inducer molecule is released from the infected bacterium and is detected by genetically modified bacterial bioreporter cells designed to emit bioluminescence upon stimulation by the inducer. Autoamplification of the bioluminescent signal permits detection of low levels of bacteria without sample enrichment. Also disclosed are methods of detection for select bacteria, and kits for detection of select bacteria based on the described technology.

22. Breathable non-woven composite viral penetration barrier fabric and fabrication process. Langley, J. D., Hinkle, B. S. (1998). Patent number 5,728,451. A breathable non-woven fabric having barrier capabilities to biological liquids comprised of at least one non-woven layer bonded to at least one surface of a thermoplastic microporous film, the non-woven composite fabric providing a barrier to passage: (a) of biological liquid when the composite fabric is subjected to contact with synthetic blood under the dictates of testing procedure ASTM ES21-92; and (b) to viral penetration when the composite fabric is subject to contact with fX174 bacteriophage suspension at a titer of 10³ PFU/mL for 5 minutes with no applied pressure, 1 minute at 13.8 kPa (2.0 PSIG), and 54 minutes with no applied pressure while maintaining a moisture of vapor transmission rate of greater than about 450 grams per square meter for 24 hours at about 75º F. and about 65% relative humidity, the non-woven composite fabric which has been thermally bonded by unwinding and contacting at least one continuous thermoplastic non-woven web to at least one side of a continuous thermoplastic microporous film, continuously transporting said contacted webs and film through a thermal bonding zone and thermally bonding the webs and film at multiple spaced-apart locations, said bonding having a dwell time sufficient to thermally bond said composite while avoiding burn-through degradation of the film and webs.

23. Cheese making with bacteriophage resistant bacteria. Hicks, C. L. (2001). Patent number 6,297,042 . A method is provided for reducing or preventing bacteriophage attack on bacteria used in a cheese making process. The method includes (a) treating a blocker peptide precursor with a protease enzyme that hydrolyzes the precursor to produce blocker peptides; (b) collecting the blocker peptides so produced; (c) formulating a starter media with the blocker peptides; (d) growing bulk cultures of cheese making bacteria in the inoculated starter media; and (e) adding bacteria grown in the inoculated starter media to a fermentation medium for producing cheese. The present invention also includes a method of making cheese and cheese produced by the method.

24. Composition for treating dental caries caused by Streptococcus mutans. Fischetti, V., Loomis, L. (2002). Patent number 6,399,098. The present invention discloses a method for treating baterial dental caries caused by Streptococcus mutans, comprising administering a composition comprising an effective amount of at least one lytic enzyme produced by a bacteriophage specific for Streptococcus mutans, with the lytic enzyme having the ability to exclusively digest a cell wall of the Streptococcus mutans infecting all or part of a mouth or teeth, and a toothpaste for delivering the enzyme to the mouth and teeth.

25. Composition for treatment of a bacterial infection of the digestive tract. Fischetti, V., Loomis, L. (2002). Patent number 6,399,097. An enteric coated pill for treating bacterial infections of the digestive tract, wherein the bacteria to be treated are selected from the group consisting of Listeria, Salmonella, E. coli, Campylobacter and combinations thereof, said pill comprising an effective amount of at least one lytic enzyme genetically coded by a bacteriophage specific for said bacteria of the digestive tract, whereby said enzyme has the ability to digest the cell wall of said bacteria; and a carrier for said enzyme.

26. Composition for treatment of a bacterial infection of an upper respiratory tract. Fischetti, V., Loomis, L. (2003). Patent number 6,423,299. An aerosol composition for treating bacterial infections of the respiratory tract by delivering said aerosol to the mouth, throat or nasal passage, wherein the bacteria to be treated is selected from the group consisting of Streptococcus pneumoniae, Haemophilus influenzae, Streptococcus Group A, and combinations thereof, said aerosol comprising an effective amount of at least one lytic enzyme genetically coded by a bacteriophage specific for a specific said bacteria of the respiratory tract, whereby said at least one lytic enzyme has the ability to digest the cell wall of said specific bacteria; and a carrier for delivering said enzyme.

27. Composition incorporating bacterial phage associated lysing enzymes for treating dermatological infections. Fischetti, V., Loomis, L. (2001). Patent number 6,277,399. A composition for treatment of bacterial infections of burns and wounds of the skin comprises an effective amount of at least one lytic enzyme produced by a bacteria infected with a bacteriophage specific for said bacteria, and a carrier for delivering said at least one lytic enzyme to the skin. The carrier may be, but is not limited to, a liquid solution applied to a bandage.

28. Compositions and methods for phage resistance in dairy fermentations. Broadbent, J. R., Oberg, C. J., Caldwell, S. (1997). Patent number 5,677,166. A latococcal- and streptococcal-phage-resistant starter culture for fermenting milk comprises a food-grade bacterium from the genera Pediococcus, Leuconostoc, Lactococcus, Streptococcus, or Lactobacillus transformed with a genetic element containing genes for a lactose fermentation phenotype. A method of making a lactococcal-phage-resistant starter culture comprises transforming a non-lactose fermenting, food-grade bacterium with a genetic element carrying determinants for a lactose fermentation phenotype. A method of making cheese with lactococcal-phage-resistant starter culture is also disclosed.

29. Compositions containing bacteriophages and methods of using bacteriophages to treat infections. Ghanbari, H. A., Averback, P. (2000). Patent number 6,121,036. Purified, host-specific, non-toxic, wide host range and virulent bacteriophage preparations that are effective in killing bacterial organisms in vivo are disclosed. Also disclosed are compositions containing these bacteriophages, methods of making the bacteriophage preparations and methods of treating bacterial infections using the compositions. Methods of treating bacterial infections using the compositions containing the bacteriophages in combination with conventional antibiotics also are disclosed.

30. Conferred susceptibility to lambda phage in non-coliform procaryotic hosts. Ludwig, R. A., de Vries, G. E. (1988). Patent number 4,784,952. Vectors suitable for effecting expression of lamB protein in a desired procaryotic host, and methods for their construction, are disclosed. Transformation with these vectors results in the ability of the procaryotic host to sustain infection by lambda phage.

31. Detection of Listeria by means of recombinant bacteriophages. Scherer, S., Loessner, M. (1998). Patent number 5,824,468. The invention relates to a detection procedure for bacteria of the genus Listeria, comprising the steps: (a) provision of a DNA vector prepared by means of recombination techniques, comprising a genetic system comprising DNA which encodes the expression of one or more proteins, the proteins not being a gene product of bacteria of the genus Listeria and it being possible to determine the presence of the proteins by a detection reaction, and the DNA vector infecting the bacteria of the genus Listeria and it being possible in this way to transfer the genetic system to the bacteria; (b) mixing of the sample with said DNA vector under conditions which allow an infection of bacteria of the genus Listeria by the DNA vector; (c) expression of the detectable proteins in the bacteria of the genus Listeria; (d) detection of the detectable proteins, the presence of bacteria of the genus Listeria being detected, and to recombinant DNA vectors and reagent compositions suitable for this detection procedure.

32. Device and method for phage-based antibiotic susceptibility testing. Cottingham, H. V. (1999). Patent number 5,858,693. A phage-based antibiotic susceptibility test is carried out by maintaining a patient sample in a sealed sample well during addition of the phage and Luciferin substrate used in the test, in order to prevent contamination of the laboratory environment. The phage is adhered in dried form to a metal carrier disk which is retained beneath the cap of the sealed sample well by means of an external magnet, and is mixed with the patient sample by removing the external magnet and allowing the carrier disk to fall to the bottom of the sample well. The Luciferin substrate is adhered to the underside of the cap and is mixed with the patient sample by shaking or inverting the sealed sample well after the metal carrier disk has separated from the underside of the cap. A row of connected sample wells and caps may be employed to allow the same patient sample to be tested with multiple antibiotics.

33. DNA encoding phage abortive infection protein from Lactococcus lactis and method of use thereof. Moineau, S., Holler, B. J., Emond, E., Vandenbergh, P. A., Vedamuthu, E. R., Kondo, J. K. (1999). Patent number 5,928,688 (see also 5,910,571) . DNA encoding phage resistance protein which aborts infection by the phage, designated as AbiE. The DNA which is contained in a Lactococcus lactis deposited as NRRL-B-21443 and described in SEQ ID NO:1, is incorporated into a bacterium to encode the AbiE and provide phage resistance. Lactococcus and other bacteria encoding the AbiE are useful in industrial fermentations wherein phage are a problem.

34. DNA encoding phage resistance protein. Moineau, S., Emond, E., Walker, S., Vedamuthu, E. R., Kondo, J. K. (1999). Patent number 5,994,118. A novel protein (Abi900, 183 amino acids) and its gene were isolated from a 11-kb natural plasmid (pSRQ900) of Lactococcus lactis. When pSRQ900 is introduced into dairy starter cultures, the Abi900 protein confers strong resistance to bacteriophage infection.

35. DNA encodoing phage abortive infection protein from Lactococcus lactis and method of use thereof. Moineau, S., Holler, B. J., Vandenbergh, P. A., Vedamuthu, E. R., Kondo, J. K. (1998). Patent number 5,814,499. DNA encoding phage resistance protein which aborts infection by the phage, designated as AbiE. The DNA which is contained in a Lactococcus lactis deposited as NRRL-B-21443 and described in SEQ ID NO: 1, is incorporated into a bacterium to encode the AbiE and provide phage resistance. Lactococcus and other bacteria encoding the AbiE are useful in industrial fermentations wherein phage are a problem.

36. DNA fragments coding for a bacteriophage-resistant mechanism. Chopin, M.-C., Cluzel, P. J. (1997). Patent number 5,629,182. The invention relates to DNA fragments encoding an Abi-type mechanism of resistance to bacteriophages, which fragments are capable of being obtained by cloning of chromosomal or plasmid DNA of a bacteriophage-resistant lactic acid bacterial strain, as well as to a polypeptide involved in an Abi-type resistance mechanism, encoded by one of the said fragments. The invention also encompasses recombinant vectors and transformed bacterial strains comprising the said DNA fragments.

37. Enzyme for phage resistance. Moineau, S., Walker, S. A., Vedamuthu, E. R., Vandenbergh, P. A. (1999). Patent number 5,972,673. An isolated DNA of a Lactococcus lactis showing a SEQ ID NO:1 encoding a restriction and two modification enzymes (R/M SEQ ID NO: 2, 3 and 4). The isolated DNA is used to transform sensitive dairy cultures, such as Lactococcus lactis and Streptococcus thermophilus, to provide phage resistance. Escherichia coli can be used to produce endonucleases.

38. Externalization of products of bacteria. Auerbach, J. I., Rosenberg, M. (1987). Patent number 4,637,980. A bacterial product is made by transforming a temperature sensitive lysogen with a DNA molecule which codes, directly or indirectly, for the product, culturing the transformant under permissive conditions and externalizing the product by raising the temperature to induce phage encoded functions.

39. Filtration medium. Degen, P. J., Bilich, M. H., Staff, T. A., Gerringer, J., Salinaro, R. F. (1998). Patent number 5,788,862. The present invention provides a filtration medium comprising an ultrafiltration membrane and a monomer surface coating thereon of an acrylic or methacrylic acid monomer having alcohol functional groups, wherein the filtration medium after having been fully dried is characterized by having a titer reduction of at least about 10³ with respect to PP7 bacteriophage and a critical wetting surface tension of at least about 70 mN/m. The filtration medium preferably further comprises a fibrous nonwoven web embedded in the membrane. The present invention also provides a method of filtering a fluid through the present inventive filtration medium, as well as a method of preparing such a filtration medium.

40. Genetically engineered reporter bacteria for the detection of bacteriophage. Rees, C. E. D., Rostas-Mulligan, K., Park, S. F., Denyer, S. P., Anderson, G. S., Stewart, B., Jassim, S. A. A. (1998). Patent number 5,723,330. A method for testing for target bacteria involves adding bacteriophage to a sample to infect the bacteria in the sample; killing extracellular bacteriophage without killing phage-infected bacteria; amplifying bacteriophage remaining in the sample; and causing the bacteriophage to infect reporter bacteria and thereby produce an observable signal. The reporter bacteria are genetically engineered to have an indicator gene which on expression gives rise to a detectable signal, wherein expression of the indicator gene is initiated on bacteriophage infection of the bacteria.

41. Inviable phages, their production and DNA thereof. Mattson, T. L., Epstein, R. (1998). Patent number 5,834,291 (see also 5,559,018). Inviable T4 phage-like particles capable of directing the expression of large non-T4 DNA fragments from T4 expression control sequences are produced. Thus, E. coli harboring pBR322 derivatives containing cloned T4 gene 23 DNA sequences were infected with T4 phage carrying a deletion of the denB gene. Homology-dependent recombination results in the production of inviable phage-like particles containing DNA molecules composed of multiple, tandemly repeated copies of entire plasmid molecules covalently linked to single copies of normal phage genes. The yield of these inviable particles, intially low, was increased by means of a reiterated infection process that involves the use of a cloned T4 origin of replication. When T4 gene 32 expression control sequences linked in proper orientation to a DNA sequence coding for the non-T4 protein b-galactosidase were also cloned in one such pBR322 derivative (pVH773), inviable phage particles capable of directing the synthesis of enzymatically active b-galactosidase were produced. The present process is applicable to other T-even bacterio-phages.

42. Isolated DNA encoding enzyme for phage resistance. Moineau, S., Walker, S., Vedamuthu, E. R., Vandenbergh, P. A. (2003). Patent number 5,925,388 (see also 5,824,523). An isolated DNA of a Lactococcus lactis showing a SEQ ID NO:1 encoding a restriction and twp modification enzymes (R/M SEQ ID NO: 2, 3 and 4). The isolated DNA is used to transform sensitive dairy cultures, such as Lactococcus lactis and Streptococcus thermophilus, to provide phage resistance. Escherichia coli can be used to produce endonucleases.

43. Lysogenic bacteriophage isolated from acidophilium. Ward, T. W., Bruhn, D. F., Bulmer, D. K. (1992). Patent number 5,132,221. A bacteriophage identified as fAc1 capable of infecting acidophilic heterotropic bacteria (such as Acidiphilium sp.) and processes for genetically engineering acidophilic bacteria for biomining or sulfur removal from coal are disclosed. The bacteriophage is capable of growth in cells existing at pH at or below 3.0. Lytic forms of the phage introduced into areas experiencing acid drainage kill the bacteria causing such drainage. Lysogenic forms of the phase having genes for selective removal of metallic or nonmetallic elements can be introduced into acidophilic bacteria to effect removal of the desired element form ore or coal.

44. Matrices with memories and uses thereof. Nova, M. P., Senyei, A. E., Potash, H. (2003). Patent number 6,100,026 (see also 5,961,923; 6,017,496; 6,340,588). Combinations, called matrices with memories, of matrix materials that are encoded with an optically readable code are provided. The matrix materials are those that are used in as supports in solid phase chemical and biochemical syntheses, immunoassays and hybridization reactions. The matrix materials may additionally include fluophors or other luminescent moieties to produce luminescing matrices with memories. The memories include electronic and optical storage media and also include optical memories, such as bar codes and other machine-readable codes. By virtue of this combination, molecules and biological particles, such as phage and viral particles and cells, that are in proximity or in physical contact with the matrix combination can be labeled by programming the memory with identifying information and can be identified by retrieving the stored information. Combinations of matrix materials, memories, and linked molecules and biological materials are also provided. The combinations have a multiplicity of applications, including combinatorial chemistry, isolation and purification of target macromolecules, capture and detection of macromolecules for analytical purposes, selective removal of contaminants, enzymatic catalysis, cell sorting, drug delivery, chemical modification and other uses. Methods for tagging molecules, biological particles and matrix support materials, immunoassays, receptor binding assays, scintillation proximity assays, non-radioactive proximity assays, and other methods are also provided.

45. Method and buffered bulk starter media for propagation of useful bacteria. Sandine, W. E., Huggins, A. R. (1988). Patent number 4,766,076. The invention comprises a novel starter medium for the commercial propagation of acid producing bacteria, such as those used in food fermentation processes. The compositions are unique in that they contain a highly effective buffering ingredient which is a sodium, potassium, or ammonium salt or double salt of a linear aliphatic dibasic acid having from three to seven carbon atoms. The salts are present in an amount sufficient to maintain the growth medium at pH levels of about 5.0 or above during the time in which the bacteria are multiplying in the culture medium. Disodium or diammonium succinate, glutarate, or adipate are materials which have been found to be particularly effective. These may be used in combination with nutrients such as whey, whey permeate, nonfat dried milk, yeast extract, and diammonium phosphate. The addition of trace quantities of certain metals promotes the growth and activity of the acid producing bacteria. Small quantities of ferrous, manganous, or manganese ions are particularly useful. A combination of iron and manganese with the other ingredients in the media produced results better than either of these materials standing by itself. In a commercial test, the cheese produced using an inoculant based on one of the present formulations was of excellent quality. Most of the formulations containing the bibasic acid salts appear to be highly resistant to bacteriophage infection.

46. Method and compositions for use in the treatment of fireblight. Vedamuthu, E. R., Vidaver, A. K. (1988). Patent number 4,783,406 (see also 4,678,750). A method and compositions for the treatment of fireblight disease in plants are described. The compositions include a phage for Erwinia amylovora which produces fireblight and an enzyme produced by the phage which depolymerizes a polysaccharide produced by Erwinia amylovora which is the cause of the fireblight disease. Purified enzyme preparations are described.

47. Method and device for detecting bacteriophage using contrast-coloring and precipitable dyes. Wicks, J. H., Krejcarek, G. E., Williams, M. G. (2000). Patent number 6,090,541. The use of a precipitable dye and a contrast-coloring dye together enhance visualization of plaques in confluent lawns of bacteria in bacteriophage and bacteria assays. A test sample suspected of containing a bacteriophage is combined with bacteria capable of replicating the bacteriophage, and applied to a water-proof surface to form a support for bacterial growth. The support is provided with the contrast-coloring dye and precipitable dye, and nutrients and salts capable of supporting growth of the bacteria. A lawn of bacteria is formed on the support, and plaques detected on the lawn indicate presence of the bacteriophage. The plaques contain a precipitate formed by enzymatic cleavage of the precipitable dye by an enzyme of the bacterial lawn. A similar procedure is used for detecting bacteria, except that a test sample suspected of containing a bacteria is combined with bacteriophage capable of replicating in the bacteria, and plaques detected indicate presence of the bacteria. The bacteriophage and bacteria assays are carried out with a disposable device containing at least one well having a water-proof surface and a depth of about at least 5 millimeters. A hydratable material containing the contrast-coloring dye and precipitable dye is positioned on the surface. The well may contain substantially vertical sides with a removable cover resting on top of the sides.

48. Method and test kits for detection of bacteriophage. Sanders, M. F. (1999). Patent number 5,914,240 . A method for detection, identification and/or quantification of bacteriophage of bacterial host specificity for bacterial genus, species or serotype, based upon the occurrence of release of cell contents, particularly nucleotides e.g. ATP, on lysis of bacterial cell walls on incubation with bacterial host cells. When new phage particles are released at the end of the phage replication cycle nucleotide levels are measured and compared with controls. The method provides for the detection of specific phages which is faster and more sensitive than known techniques. The method is only limited by the availability of host bacteria/target phage pairings.

49. Method for detecting bacteria using bacteriophage, contrast-coloring dye and precipitable dye. Wicks, J. H., Krejcarek, G. E., Williams, M. G. (1999). Patent number 5,958,675. Bacteria are detected in a test sample by contacting the test sample with a bacteriophage that is capable of replicating in the bacteria, adding the resultant sample to a water-proof surface of a support for bacterial growth that contains a contrast-coloring dye and a precipitable dye, forming a bacterial lawn of a bacteria in which the bacteriophage can replicate on the support and detecting plaques on the bacterial lawn as an indication of the presence of the bacteria. The combination of precipitable dye and contrast-coloring dye improves visualization of plaques. A precipitate is formed in plaques by enzymatic cleavage of the precipitable dye by an enzyme of the bacterial lawn. A procedure for detecting bacteriophage is similar to that for detecting bacteria, except that a test sample suspected of containing bacteriophage is combined with bacteria in which the bacteriophage can replicate, and plaques detected indicate presence of the bacteriophage. The bacteria and bacteriophage detections are carried out with a disposable device containing at least one well having a water-proof surface and substantially vertical sides that extend at least 5 millimeters in height from the surface. A hydratable material containing the precipitable dye and the contrast-coloring dye is positioned on the water-proof surface. A removable cover rests on top of the sides of the well.

50. Method for detecting bacteria with bacteriaphage [sic]. Nakayama, , H. (2003). Patent number 6,555,312. A method for detecting a bacterium for measurement, including the steps of: allowing a bacteriophage to bind to the bacterium, the bacteriophage being capable of specifically binding to the bacterium and growing in the bacterium, whereby a gene within the bacteriophage which expresses a light-emission protein is introduced into the bacterium so that a protein is produced within the bacterium as a product of the gene; and providing an external factor in a non-invasive manner from outside of the bacterium, thereby causing only the actually-present bacterium to emit light in a specific manner.

51. Method for facilitating externalization of proteins synthesized in bacteria. Zinder, M. D., Model, P., Boeke, J. D. (1986). Patent number 4,595,658. A method of externalizing proteins from the periplasmic space of gram-negative bacteria and in particular, E. coli and its relatives, comprising utilizing bacteria which have a phage gene, coding for a protein (such as gene III protein) or a mutant bacterial gene (such as a gene coding for a membrane function) which causes perturbation of the outer bacterial membrane resulting in leakage of proteins in the periplasmic space from the cell.

52. Method for forming an array of biological particles. Raybuck, M. (1998). Patent number 5,763,170. A method for forming and using an array of, e.g., bacteria, yeast or bacteriophage for the purpose of identifying particular constituents thereof. The array is formed by directing a stream of droplets, each containing on average about 1 or a few biological particles, at spaced locations in an array on a surface, e.g., a nylon membrane or agar gel.

53. Method for identifying target bacteria. Sanders, M. F. (1999). Patent number 5,888,725. A method for detection, identification and/or quantification of target organisms of specific bacterial genus, species or serotype, based upon the occurence of release of cell contents, particularly nucleotides, e.g., ATP, on lysis of bacterial cell walls on incubation with bacteriophages (phages) specific for them. When new phase particles are released at the end of the phage replication cycle nucleotide levels are measured and compared with controls. The method provides for the detection of specific bacteria which does not require insertion of the lux gene into the phage genome yet is faster and more sensitive than known non-modified phage utilizing techniques. The method is only limited by the availability of phage types suitable for selective attack of the target bacterial to be detected and can detect a single Salmonella in a sample of milk in under 12 hours.

54. Method for producing single and/or mixed strain concentrates of bacteria. Sandine, W. E., Huggins, A. R. (1985). Patent number 4,528,269 . An improved method which differentiates or separates heterogeneous populations of fast and slow acid producing strains of bacteria by growth of the strains under closely controlled unique conditions so as to allow the selection of a colony of one or the other strains is described. Preferably a gelled, solid growth medium containing in admixture: (1) milk protein, a milk protein derivative, or a milk protein substitute; (2) an acid pH sensitive color change indicator; and, (3) a buffering agent is used. The colonies have a contrasting color within and around them because of the effect of the acid produced by the bacteria on the indicator. The growth of the bacteria is under anaerobic or near anaerobic conditions in order to achieve certainty in the colony selection for fast or slow acid production. The bacteria can also be mixed with phage which inhibit or kill the members of a heterogeneous or homogeneous population of bacteria on the medium and grown to produce phage resistant colonies. The relatively large colonies which exhibit enhanced acid production and proteolysis of the milk protein on the plating container are selected for commercial use in preparing fermented products, particularly fermented foods.

55. Method for producing mucoid and phage resistant group N streptococcus strains from non-mucoid and phage sensitive parent strains. Vedamuthu, E. R. (1990). Patent number 4,918,014 (see also 4,874,616). A method for imparting phage resistance to phage sensitive strands of Streptococcus group N is described. The method involves transferring plasmid encoding for production of a mucoid substance (Muc⁺) into the phage sensitive strain. Even if the Muc⁺ plasmid is removed by curing at elevated temperatures the strains remain resistant to phage. The resulting resistant strains are novel and are used for fermentations, particularly milk fermentations.

56. Method of conferring bacteriophage resistance to bacteria. Hershberger, C. L., Rosteck, P. R. (1985). Patent number 4,530,904. A novel method for protecting a bacterium from a naturally occurring bacteriophage and the cloning vectors and transformants for carrying out the aforementioned method are disclosed.

57. Method of detecting compounds utilizing genetically modified lambdoid bacteriophage. Ray, B. L., Lin, E. C. C., Crea, R. (1997). Patent number 5,650,267. Disclosed is an infective lambdoid bacteriophage which includes a protein construct comprising a genetically modified major tail protein truncated at its carboxy terminus, and a target molecule peptide bonded to the carboxy terminus of the tail protein. Also disclosed are nucleic acids encoding the construct and methods of detecting a molecule-of-interest in a solution and of detecting a cell which produces a molecule-of-interest.

58. Method of detecting a pathogen using a virus. Cherwonogrodzky, J. W., Lotfali, K. (2002). Patent number 6,436,652 (see also 6,355,445). A bacteriophage linked to an enzyme can replace an antibody in a system for detecting the presence of a bacteria in a sample. Specifically Brucella abortus (a pathogen which causes brucellosis in cattle) can be detected using Brucella bacteriophage for the virus, urease for the enzyme linked to the bacteriophage, m-maleimidobenzoyl-N-hydrosysuccimide ester as a coupling reagent, sera from mice immunized with Brucella bacteriophage for a detector antibody, urease conjugated to anti-mouse sheep antibody for an indicator, and urea with bromcresol purple as the substrate. The materials can be used in indirect (sandwich) or direct enzyme-linked viral assays (ELVirA).

59. Method of eliminating genetic routes for bacteriophage evolution and products produced thereby. Klaenhammer, T. R., Moineau, S. (1997). Patent number 5,618,723 (see also 5,580,725). A process of identifying and disrupting bacterial DNA sequences that contribute to the evolution of new lytic bacteriophages is described. Vectors and recombinant bacteria for use in producing fermentative starter cultures and culture resistant to the appearance of new phages, and methods of producing such vectors and recombinant bacteria, are described.

60. Method of making cheese using viral enzymes. Gasson, M. J. (1994). Patent number 5,360,617. The lysin from a Lactococcus (preferably prolate-headed) bacteriophage is used to lyse bacterial starter cultures during cheese-making. Such bacteriophage include fvML3. In addition, the fvML3 lysin has been characterized and a coding sequence for it has been cloned.

61. Method of preparing cheese starter media. Reddy, M. S. (1986). Patent number 4,621,058. Low cost, readily dispersible, phage-resistant cheese starter media are described which include milk-derived nutrients (e.g., nonfat milk and whey) along with a minor proportion of preferably free or unbound lecithin. The media also may advantageously include sodium tetraphosphate which assists in the dispersion of whey solids. The media of the invention can be used at significantly lower levels as compared with nonfat dry milk solids (e.g. 7 percent versus 12 percent), while nevertheless obtaining essentially equivalent results in terms of culture growth and final culture properties. A method of producing the media is also disclosed, involving liquid preblending of phosphates and lecithin, followed by addition thereof to milk-derived nutrients and reaction of the phosphates to tie up free calcium ion. Preferably, reaction is carried out for about 1 to 12 hours while agitating. The final step involves drying of the mixture to yield a substantially homogeneous, reconstitutable powder. In other cases the phosphate-lecithin preblend can be dried for later addition to milk-derived nutrients to produce a final starter medium.

62. Method of preparing food and composition for protecting microorganisms used in the preparation of food. Lembke, A., Deininger, R., Lembke, J. (1989). Patent number 4,834,987. In the preparation of food with the aid of microorganisms, the latter are directly protected against viral or phage attack by the addition of formic acid or esters of formic acid or salts of formic acid and/or tetrahydrofolic acid. Furthermore, an indirect protection by inactivating the bacterial viruses in the environment is described.

63. Method of recovering bacteriophage. DeBonville, D. A., Logan, K. A. (1993). Patent number 5,204,257. A method of recovering nucleic acid-containing particles from a liquid medium by contacting the liquid medium containing the particles with a mixture of hydroxylated silica beads and a salt solution to bind the nucleic acid-containing particles, centrifuging the mixture to pellet the bound particles, and separating the pellet from the liquid.

64. Method to detect bacteria. Wilson, S. M. (2002). Patent number 6,461,833 (see also 5,985,596). The present invention relates to a method for enhancing the time of response of an assay for a first bacterium, wherein: a) the first bacterium is exposed to infection by phage particles to which the first bacterium is permissive; b) the infected bacterium is treated to inactivate exogenous phage particles; c) the treated bacterium is cultivated in the presence of a second bacterium which is permissive to infection by the phage or its replicand and which has a doubling rate greater than the effective doubling rate of the first bacterium; and d) assessing the extent of plaque formation and/or of second bacterium growth in the cultivated second bacterium cells. The method can be used to assess the presence of first bacterium in a sample, notably where the first bacterium is a slow growing bacterium, such as Mycobacterium tuberculosis, where the method enables an operator to detect the presence of low amounts of the bacterium in sample within days instead of weeks as required by conventional cultivation techniques. The invention can also be used to assess the effect of a drug or other treatment on a bacterium or on a virus. The invention also provides a diagnostic kit for use in the method of the invention.

65. Methods for rapid microbial detection. Rees, C. E. D., Rostas-Mulligan, K., Park, S. F., Denyer, S. P., Stewart, G. S. A. B., Jassim, S. A. A. (1996). Patent number 5,498,525. A method of testing for target bacteria involves adding bacteriophage to a sample to infect the bacteria in the sample; killing extracellular bacteriophage without at the same time killing phage-infected bacteria; amplifying bacteriophage remaining in the sample; and causing the bacteriophage to infect reporter bacteria and thereby produce an observable signal. The reporter bacteria are genetically engineered to have an indicator gene which on expression gives rise to a detectable signal, wherein expression of the indicator gene is initiated on bacteriophage infection of the bacteria.

66. Methods of detection utilizing modified bacteriophage. Li, M. (2001). Patent number 6,190,856. Viruses expressing ligands on their surfaces are used as a detection means for the related polypeptide which binds the ligand. Multiple copies of the ligand can be expressed on the viral surface. These viruses may be used to detect polypeptides, cells, receptors and channel proteins.

67. Mycobacteriophage DSGA specific for the Mycobacterium tuberculosis complex. Pearson, R. E., Dickson, J. A., Hamilton, P. T., Little, M. C., Beyer, Jr. W. F. (1995). Patent number 5,476,768. Mycobacteriophage DS6A has been characterized and found to specifically infect all species of the TB complex, without any detectable infection of mycobacteria species other than those of the TB complex. DNA sequence analysis revealed several potential open reading frames, including one encoding a protein analogous to gp37 of mycobacteriophage L5 and a second encoding a protein with significant homology to the S. coelicolor DNA polymerase b subunit. Based on the DNA sequence analysis, cloning sites can be identified for insertion of reporter genes, making DS6A useful as a reporter phage for specific detection and identification of species of the TB complex.

68. Mycobacteriophage specific for the Mycobacterium tuberculosis complex. Pearson, R. E., Dickson, J. A., Hamilton, P. T., Little, M. C., Beyer, Jr. W. F. (1997). Patent number 5,612,182 (see also 5,582,969). Mycobacteriophage DS6A has been characterized and found to specifically infect all species of the TB complex, without any detectable infection of mycobacteria species other than those of the TB complex. DNA sequence analysis revealed several potential open reading frames, including one encoding a protein analogous to gp37 of mycobacteriophage L5 and a second encoding a protein with significant homology to the S. coelicolor DNA polymerase b subunit. Based on the DNA sequence analysis, cloning sites can be identified for insertion of reporter genes, making DS6A useful as a reporter phage for specific detection and identification of species of the TB complex.

69. Non-isotopic substrate assay employing bacteriolysis products. Young, D. M. (1978). Patent number 4,104,126. Substrates such as haptens and antigens, and those for receptor proteins and native circulating binding proteins are assayed by determining bacteriolysis products occasioned by bacteriophage infection of host cells, in a modification of the "chemically modified bacteriophage assay." Thus, a substrate such as an antigen is conjugated with bacteriophage and the conjugate competes with antigen in the specimen under assay for a limited number of binding sites on antibody. Phage conjugate surviving antibody inactivation is quantified by determining intracellular constituents of host bacteria subsequently infected by the bacteriophage remaining viable, which latter can be related to the levels of antigen originally present in the specimen. A preferred embodiment involves colorimetric assay for beta galactosidase freed by phage lysis of E. coli. Generally, the method is of sensitivity comparable to that of radioimmunoassay, but is attended by substantial advantages not common to the latter technique. The method is far less cumbersome than the plaque-containing techniques hitherto employed in bacteriophage assays.

70. Novel bacteriophage and method for preparing same. Nakano, E., Saito, N., Fukushima, D. (1982). Patent number 4,332,897. A novel bacteriophage whose DNA molecule has endonuclease-sensitivity only in the DNA region carrying genetic information for the production of phage coat proteins can be obtained by isolating an endonuclease-resistant mutant from one of the lambdoid bacteriophages and mating the resulting bacteriophage with a lambdoid phage having endonuclease-sensitivity in the DNA region carrying genetic information for the production of coat proteins.

71. Nucleic acid sequence and plasmids comprising at least one phage resistance mechanism, bacteria in which they are present, and their use. Prevots, F., Remy, E., Ritzenthaler, P. (1997). Patent number 5,658,770. The invention relates to a DNA sequence of about 1.9 kb comprising at least one phage resistance mechanism, said sequence being obtained from the HindIII--HindIII DNA sequence of 3.3 kb contained in the strain Lactococcus lactis ssp lactis, deposited in the CNCM under no. I-945, by the PCR method.

72. Nucleic acid sequences and plasmids comprising at least one phage resistance mechanism, bacteria containing them and their use. Prevots, F., Tolou, S., Daloyau, M. (1999). Patent number 5,955,332 (see also 5,712,150). The invention relates to polynucleotides of 1345 bp and 3704 bp and the like, which comprise at least one phage resistance mechanism and obtainable from the total DNA contained in the Lactococcus lactis ssp cremoris strain deposited in the CNCM under No. I-941.

73. Parenteral use of bacterial phage associated lysing enzymes for the therapeutic treatment of bacterial infections. Fischetti, V., Loomis, L. (2001). Patent number 6,264,945 . The present invention discloses a method and composition for the treatment of bacterial infections by the parenteral introduction of at least one lytic enzyme produced by a bacteria infected with a bacteriophage specific for that bacteria and an appropriate carrier for delivering the lytic enzyme into a patient. The injection can be done intramuscularly, subcutaneously, or intravenously.

74. Phage-dependent super-production of biologically active protein and peptides. Kordyum, V. A., Chernykh, S. I., Slavchenko, I. Y., Vozianov, O. F. (2001). Patent number 6,268,178. This invention relates to a method for enhancing the production of biologically active proteins and peptides in bacterial cells by infecting bacterial cells of the producer strain, which contain a plasmid with one or more targeted genes, with bacteriophage .lambda. with or without the targeted gene(s). The phage increases synthesis of the targeted protein and induces lysis of the producer strain cells. Super-production is achieved by cultivating the producer strain cells under culture conditions that delay lytic development of the phage. The biologically active proteins and peptides subsequently accumulate in a soluble form in the culture medium as the cells of the producer strain are lysed by the phage.

75. Phage-resistant streptococcus. Mollet, B., Pridmore, D., Zwahlen, M. C. (1998). Patent number 5,766,904. DNA fragment of phages which are virulent towards a Streptococcus, capable of conferring on a Streptococcus containing it resistance to at least one phage, especially a fragment homologous or hybridizing to the 3.6 kb HindIII fragment present in the plasmid CNCM I-1588 or the 6.5 kb EcoRV fragment present in the plasmid CNCM I-1589. Process for making a Streptococcus resistant to at least one phage, by cloning into a vector a DNA fragment of a phage which is virulent towards a Streptococcus, capable of conferring on a Streptococcus resistance to at least one phage and introducing the vector into a Streptococcus.

76. Phage bonded to a nuclear location signal. Nakanishi, M., Nagoshi, E., Akuta, T., Takeda, K., Hasegawa, M. (2001). Patent number 6,235,521 . A l phage with a nuclear localization signal has been obtained by constructing a vector capable of expressing a fused protein between a gpD protein constituting the head of a l phage and a nuclear localization signal sequence, transforming Escherichia coli with this vector, and propagating a mutant l phage which cannot express the gpD protein in E. coli in this transformant. It has been confirmed that the resulting l phage is capable of packaging l phage DNAs of 80% and 100% genome sizes. After further confirming that the nuclear localization signal exposed on the outside of the head of this phage, this phage has been microinjected into cells to analyze its nuclear localization activity. Thus, it has been clarified that this phage has a nuclear localization activity.

77. Phage defense rotation strategy. Klaenhammer, T. R., Sing, W. D., Hill, C. J. (1997). Patent number 5,593,885. A phage defense rotation strategy for use in the successive fermentations of a substrate in a fermentation plant is disclosed. The strategy comprises (a) fermenting substrate with a first bacterial culture comprising a bacterial strain capable of fermenting the substrate and, preferably, carrying a first phage defense mechanism; and then (b) fermenting the substrate with a second bacterial culture comprising a second bacterial strain isogenic with the first bacterial strain, wherein the second strain carries a second phage defense mechanism different from the first phage defense mechanism. Also disclosed is a mixed bacterial culture capable of fermenting a substrate. The mixed culture comprises (a) a first bacterial strain carrying a first phage defense mechanism; and (b) a second bacterial strain isogenic with the first strain, wherein the second strain carries a phage defense mechanism different from the phage defense mechanism carried by the first strain.

78. Phage detection. LaBelle, G. G., Staehler, G. E. (1980). Patent number 4,218,534. To select a blend of strains not susceptible to the current bacteriophage in the cheesemaking plant the cheesemaker inoculates each of the test tubes in the kit with filtered when obtained from current production. Each tube contains a genetically distinct starter culture strain or a culture blend in a sterile milk medium and contains a dye which will change color in the desired pH range. After incubation for ten hours the cultures resistant to the prevailing phage will exhibit the desired color change and will have developed a firm curd. A starter culture now known to be resistant to the prevailing phage can now be selected. Tests show success closely approaching 100% as opposed to 96% (or less) with the traditional rotation method of selecting culture blends.

79. Phage with nuclear localization signal. Nakanishi, M., Nagoshi, E., Akuta, T., Takeda, K., Hasegawa, M. (2001). Patent number 6,300,120. A l phage with a nuclear localization signal has been obtained by constructing a vector capable of expressing a fused protein between a gpD protein constituting the head of a l phage and a nuclear localization signal sequence, transforming Escherichia coli with this vector, and propagating a mutant l phage which cannot express the gpD protein in E. coli in this transformant. It has been confirmed that the resulting l phage is capable of packaging l phage DNAs of 80% and 100% genome sizes. After further confirming that the nuclear localization signal exposed on the outside of the head of this phage, this phage has been microinjected into cells to analyze its nuclear localization activity. Thus, it has been clarified that this phage has a nuclear localization activity.

80. Polyvinylidene fluoride membrane and method for removing viruses from solutions. Degen, P. J., Bilich, M. H., Staff, T. A., Gerringer, J., Salinaro, R. F. (1998). Patent number 5,736,051. The present invention provides an isotropic, skinless, porous, polyvinylidene fluoride membrane having a K_UF of at least about 15 psi (103 kPa), and preferably below about 50 psi (345 kPa), when tested using liquid pairs having an interfacial tension of about 4 dynes/cm (4 mN/m). The present inventive membrane preferably has a titer reduction of at least about 10⁸ against T₁ bacteriophage, more preferably also against PR772 coliphage, and even more preferably also against PP7 bacteriophage. The present inventive membrane can have a thickness of about 20 mil (500 mm) or less and even as low as about 5 mil (125 mm) or less. The present invention also provides a method of preparing such a membrane by providing a casting solution comprising polyvinylidene fluoride and a solvent therefor, heating the casting solution to a uniform temperature of about 57º C. to about 60º C., spreading the casting solution onto a substrate to form a film, quenching the film in a quench bath so as to form a porous membrane, and washing and drying the porous membrane.

81. Process for retarding bacterial growth in cheese. Day, C. A., Holton, B. W. (1991). Patent number 5,006,347. The present invention discloses the use of bacteriophages for controlling unwanted fermentation of cheese by bacteria.

82. Production of phage and phage-associated lysin. Yang, H.-H., Hiu, S. F., Harris, J. L. (1989). Patent number 4,859,597. A method for producing a lysin-free phage inoculum, which comprises: (a) inoculating a growing Streptococcal culture with phage, (b) incubating the culture for a plurality of lytic cycles of phage until the cells are completely lysed to obtain a lysate, and (c) removing cell debris and free lysin from the lysate to form a lysin-free phage suitable for use as an inoculum. A method for producing lysin is also disclosed.

83. Prophylactic and theraputic treatment of group A streptococcal infection. Fischetti, V., Loomis, L. (1999). Patent number 5,985,271. The present invention relates to an oral delivery system containing a group c streptococcal phage associated lysin enzyme for the prophylactic and therapeutic treatment of Streptococcal A throat infections, commonly known as strep throat

84. Protection of microorganisms against bacteriophage attacks. Wolf, E., Lembke, A., Deininger, R. (1983). Patent number 4,409,245. Living cultures of microorganisms used in the preparation of foodstuffs by microbiological processing are protected against attack by bacteriophage viruses by the addition thereto of terpene. The terpene is added in an amount which is effective to obtain viricidal activity but ineffective to cause toxic effects on the microorganisms. The terpene is one obtainable from aromatic plants by steam distillation. Terpenes or mixtures of terpenes which have proved suitable are those obtained from black pepper oil, cinnamon flower oil, cardamon oil, linallyl acetate, cinnamic aldehyde, safrol, carvon and cis/ trans citral, used individually or mixed together. They may added dissolved in a carrier such as 1,2-propanediol. The terpenes demonstrate a viricidal activity in a concentration which is one or more powers of ten lower than the concentration at which the terpenes have toxic effects on the microorganisms.

85. pTN1060, a conjugal plasmid and derivatives thereof that confer phage resistance to group N streptococci. Klaenhammer, T. R., Sanozky-Dawes, R. B. (1989). Patent number 4,883,756. The present invention relates to the plasmid pTN1060 and derivatives thereof which confer phage restriction and modification activity to group N streptococci. The invention further relates to microorganisms containing pTN1060 or a derivative thereof and to starter cultures containing the microorganisms.

86. PTR2030, a conjugal plasmid and derivatives thereof that confer phage resistance to group N streptococci. Klaenhammer, T. R., Sanozky, R. B., Steenson, L. R. (1992). Patent number 5,139,950 (see also 4,931,396). The present invention relates to the plasmid pTR2030 and derivatives thereof which confer phage resistance to group N streptococci. The invention further relates to microorganisms containing pTR2030 or a derivative thereof and to starter cultures containing the microorganisms.

87. Quantitative detection of specific nucleic acid sequences using lambdoid bacteriophages linked by oligonucleotides to solid support. Ray, B. L., Lin, E. C. C. (2003). Patent number 5,679,510. The present invention provided compositions, methods and kits for detection and quantitation of pathogenic organisms. The composition of the invention is an oligonucleotide probe comprising a bacteriophage covalently linked to one site on an oligonucleotide probe complementary to a conserved region of a pathogenic organism. At a second site, the oligonucleotide probe is linked to a matrix. The oligonucleotide probe contains a region complementary to one strand of a restriciton endonuclease recognition site or an oligoribonucleotide moiety. The number of pathogenic organisms present in a biological fluid sample may be quantitated in accordance with the method of the invention by combining the composition of the invention with the sample, allowing hybridization to occur. Hybridization generates a DNA-RNA hybrid, and by adding the appropriate nucleolytic enzyme capable of cleaving DNA-RNA hybrids; bacteriophage will be released for measurement. The kit of the invention provides components which allow the method of the invention to be performed.

88. Rapid identification of environmental bacillus. Kiel, J. L., Alls, J. L., Weber, R. A., Parker, J. E. (1992). Patent number 5,156,971. A diagnostic test for environmental bacillus which comprises the steps of inoculating an agar growth medium comprising a nitrate source, luminol and 3-amino-L-tyrosine (3AT) with the sample, incubating the inoculated medium and determining the presence of the bacillus. The novel medium preferably comprises potassium nitrate, luminol, 3-amino-L-tyrosine and trypticase soy agar. Antibiotics and/or a specific bacteriophage may be added to the medium surface in localized areas to show specific bacterial lysis for identification. The novel medium and the methods of this invention are suitable for the identification of B. anthracis.

89. Rapid Identification of Escherichia coli O157:H7. Chang, T. C., Chen, S., Ding, H. C. (2001). Patent number 6,210,911. A method of determining whether a test microorganism is a known microorganism, involving use of an agent that specifically affects the growth of the known microorganism. The invention also features a method of identifying E. coli O157:H7 that are based on the following criteria: a test microorganism is E. coli O157:H7 if the microorganism is (i) E. coli, (ii) incapable of fermenting sorbitol, and (iii) susceptible to infection by AR1 phage.

90. Rapid identification of microorganisms. Chang, T. C., Chen, S., Ding, H. C. (2002). Patent number 6,428,976. A method of determining whether a test microorganism is a known microorganism, involving use of an agent that specifically affects the growth of the known microorganism. The invention also features a method of identifying E. coli O157:H7 that are based on following criteria: a test microorganism is E. coli O157:H7 if the microorganism is (i) E. coli, (ii) incapable of fermenting sorbitol, and (iii) susceptible to infection by AR1 phage.

91. Recombinant phages. Mardh, S. (2002). Patent number 6,497,874. The present invention relates to bacteriophages for use in the treatment or prophylaxis of bacterial infections, especially mucosal bacterial infections such as Heliobacter pylori infections, in particular, it relates to modified filamentous bacteriophages, e.g. M13 phages, for such use, which bacteriophages present at the surface a recombinant protein comprising (i) a first component derived from a bacteriophage surface protein; and (ii) a second component comprising variable region sequences of an antibody to provide a bacterial antigen binding site, said second component rendering said bacteriophage capable of binding to and thereby inhibiting growth of bacterial cells involved in the etiology of said infection.

92. Remotely programmable matrices with memories. Nova, M. P., Senyei, A. E. (2002). Patent number 6,416,714 (see also 5,741,462; 5,751,629; 5,874,214; 5,925,562; 6,025,129; 6,331,273; 6,352,854). Combinations, called matrices with memories, of matrix materials with remotely addressable or remotely programmable recording devices that contain at least one data storage unit are provided. The matrix materials are those that are used in as supports in solid phase chemical and biochemical syntheses, immunoassays and hybridization reactions. The data storage units are preferably non-volatile antifuse memories. By virtue of this combination, molecules and biological particles, such as phage and viral particles and cells, that are in proximity or in physical contact with the matrix combination can be labeled by programming the memory with identifying information and can be identified by retrieving the stored information. Combinations of matrix materials, memories, and linked molecules and biological materials are also provided. The combinations have a multiplicity of applications, including combinatorial chemistry, isolation and purification of target macromolecules, capture and detection of macromolecules for analytical purposes, selective removal of contaminants, enzymatic catalysis, chemical modification and other uses.

93. RNA bacteriophage-based delivery system. Stockley, P. G., Mastico, R. A. (2000). Patent number 6,159,728. A delivery system, especially for delivery to targeted sites in the human or animal body, comprises capsids of the coat protein amino acid sequence of phage MS-2 or related phage, or a modification thereof which retains capsid-forming capability, and at least some of the capsids enclosing a moiety foreign to the genome of MS-2 or related phage.

94. Super fast tuberculosis diagnosis and sensitivity testing method. Ledley, R. S. (1999). Patent number 5,922,282. Very rapid diagnosis and sensitivity testing can significantly stem the growing Tuberculosis epidemic in the United States, caused by susceptible AIDs patients and the occurrence of antibiotic resistant mycobacilli. Thus I have invented an automated computerized microscope, the ATBD unit, and slide module to diagnose and test patient's sputum by examining individual living mycobacteria from the patient sample with no culturing required. The diagnosis and sensitivity testing is accomplished in minutes or hours, instead of the current weeks to months. The system inserts a plasmid, specific for M. tuberculosis, carrying the luciferase gene into the mycobacteria by improved electroporesis on the slide. Luminescence indicates tuberculosis. Then the mycobacteria are bathed in antibiotics, and if the luminescence is not turned off, the patient's bacteria are resistant. A phage carrying the luciferase gene can also be used to infect the M.TB. Finally, the invention can be applied to any mycobacteriological infection to do diagnosis sensitivity testing even when the species is not known.

95. Therapeutic antimicrobial polypeptides, their use and methods for preparation. Jaynes, J. M., Enright, F. M., White, K. L. (2001). Patent number 6,303,568. A novel class of antimicrobial agents for animal species including cecropins, attacins, lysozymes, phage derived polypeptides, such as those transcribed from gene 13 of phage 22, an S protein from lambda phage, and an E protein from phage PhiXl74, as well as, synthetically derived polypeptides of similar nature. The antimicrobial agents can be used to treat microbial infections and as components of medicinal compositions. The genes encoding for such antimicrobial agents can be used to transform animal cells, especially embryonic cells. The transformed animals including such antimicrobial cells are also included.

96. Therapeutic treatment of group A streptococcal infections. Fischetti, V., Loomis, L. (2000). Patent number 6,017,528 (see also 5,997,862). The present invention relates to compositions containing Group C streptococcal phage associated lysin enzyme for the prophylactic and therapeutic treatment of Streptococcal infections, including the infection commonly known as strep throat. Methods for therapeutically and prophylactically treating such infections also are described.

97. Topical treatment of streptococcal infections. Fischetti, V., Loomis, L. (2000). Patent number 6,056,955. The present invention discloses a method and composition for the topical treatment of streptococcal infections by the use of a lysin enzyme blended with a carrier suitable for topical application to dermal tissues. The method for the treatment of dermatological streptococcal infections comprises administering a composition comprising effective amount of a therapeutic agent, with the therapeutic agent comprising a lysin enzyme produced by group C streptococcal bacteria infected with a C1 bacteriophage. The therapeutic agent can be in a pharmaceutically acceptable carrier.

98. Transducing particles and methods for their production. Gutterson, N. I., Tucker, W. T., Wolber, P. K. (1993). Patent number 5,187,061. Viable bacteria may be detected in biological samples by exposing bacterial cultures obtained from the samples to transducing particles having a known host range. Such transducing particles carry a heterologous gene capable of altering the phenotype of the bacteria in a readily detectable manner. For example, the transducing particles may carry an ice nucleation gene and the alteration of phenotype may be detected using an ice nucleation assay. By employing a panel of phage, unknown bacteria nmay be typed based on the pattern of reactivity observed. The transducing particles may be prepared by introducing a synthetic transposable element carrying the heterologous gene to a host carrying a prophage having the desired host range. After transposition, the host may be induced to a lytic cycle to release the transducing particles carrying the heterologous gene.

99. Transducing phages of Actinomycetales. Westpheling, J., Burke, J. A. (2001). Patent number 6,245,504. The present invention is directed to isolated transducing phages, methods of isolating transducing phages, and methods of using transducing phages including, for instance, transferring at least one nucleic acid fragment from a donor microbe to a recipient microbe, and producing a secondary metabolite from a microbe. The transducing phages typically have a broad host range, and transduce microbes in the Order Actinomycetales, in particular in the Family Streptomycetaceae, including Streptomyces coelicolor, Streptomyces lividans, Streptomyces venezuelae, Streptomyces avermitilis, and Saccharopolyspora erythraea. The transducing phages can be specialized transducing phages or generalized transducing phages.

100. Use of bacterial phage associated lysing enzymes for treating various illnesses. Fischetti, V., Loomis, L. (2001). Patent number 6,238,661. Compositions and methods for the prophylactic and therapeutic treatment of bacterial infections are disclosed which comprise administering to an individual an effective amount of a composition comprising an effective amount of lytic enzyme and a carrier for delivering the lytic enzyme. This method and composition can be used for the treatment of upper respiratory infections, skin infections, wounds, burns, vaginal infections, eye infections, intestinal disorders and dental problems.

101. Use of bacterial phage associated lysing enzymes for treating dermatological infections. Fischetti, V., Loomis, L. (2002). Patent number 6,432,444. A bandage for treating a bacterial infection of skin is disclosed wherein the bandage contains a composition produced by the method of obtaining an effective amount of at least one lytic enzyme genetically coded for by a specific bacteriophage specific for a bacteria infecting the skin, wherein the bacteria to be treated is selected from the group consisting of Staphylococcus, Pseudomonas, Streptococcus, and combinations thereof. This lytic enzyme is specific for and has the ability to digest a cell wall of one of the bacteria and is coded for by the same bacteriophage capable of infecting the bacteria being digested. The enzyme produced is mixed with a topical carrier.

102. Use of bacterial phage associated lysing enzymes for treating bacterial infections of the mouth and teeth. Fischetti, V., Loomis, L. (2002). Patent number 6,335,012. The present invention discloses a method for treating dental caries, comprising administering a composition comprising an effective amount of at least one lytic enzyme produced by a bacteriophage specific for said bacteria, the lytic enzyme having the ability to digest a cell wall of the bacteria infecting all or part of a mouth or teeth, and a carrier for delivering the enzyme to the mouth and teeth.

103. Use of bacterial phage associated lysing enzymes for treating streptococcal infections of the upper respiratory tract. Fischetti, V., Loomis, L. (2002). Patent number 6,326,002. The present invention discloses a method for treating Streptococcal infections of the upper respiratory tract, comprising administering an effective amount of a lysin enzyme produced by group C Streptococcal bacteria infected with a C1 bacteriophage specific for the bacteria to a mouth, throat, or nasal passage of a mammal, the method providing a concentration of from about 100 to 500,000 active enzyme units per milliliter of fluid in the environment of said nasal or oral passages.

104. Use of bacterial phage associated lysing enzymers for the prophylactic and therapeutic treatment of various illnesses. Fischetti, V., Loomis, L. (2003). Patent number 6,056,954. A method for the prophylactic and therapeutic treatment of bacterial infections is disclosed which comprises the treatment of an individual with an effective amount of a lytic enzyme composition specific for the infecting bacteria, with the lytic enzyme comprising an effective amount of lytic enzyme, wherein the lytic enzyme is in an environment having a pH which allows for activity of said lytic enzyme; and a carrier for delivering said lytic enzyme. This method, and composition can be used for the treatment of upper respiratory infections, skin infections, wounds, and burns, vaginal infections, eye infections, intestinal disorders and dental problems.

105. Use of bacteriophages for control of Escherichia coli O157. Waddell, T. E., Mazzocco, A., Pacan, J., Ahmed, R., Johnson, R., Poppe, C., Khakhria, R. (2002). Patent number 6,485,902. A method of reducing levels of E. coli O157 strains within the gastrointestinal tract of a ruminant animal using specific bacteriophage(s) is herein described. Also described is a pharmaceutical composition comprising at least one of said bacteriophages and a method for isolating or selecting bacteriophages useful in reducing E. coli O157 levels as described above.

106. Use of phage associated lytic enzymes for treating bacterial infections of the digestive tract. Fischetti, V., Loomis, L. (2001). Patent number 6,254,866. A method for treatment of bacterial infections of the digestive tract is disclosed which comprises administering a lytic enzyme specific for the infecting bacteria. The lytic enzyme is preferably in a carrier for delivering said lytic enzyme. The bacteria to be treated is selected from the group consisting of Listeria, Salmonella, E. coli, Campylobacter, and combinations thereof. The carrier for delivering at least one lytic enzyme to the digestive tract is selected from the group consisting of suppository enemas, syrups, or enteric coated pills.

107. Use of phage associated lytic enzymes for the rapid detection of bacterial contaminants. Trudil, D. (2002). Patent number 6,395,504. A method for the use of a phage associated lysing enzyme for the detecting the presence and determining the quantity of bacteria present in or on a wide variety of substances is described. The total concentration of microbes is determined by adding or incorporating a phage associated lytic agent to a disposable test system device with the luminescent reagents luciferin and luciferase, and introducing the disposable test system into a luminometer that can read the luminescence. Other systems can be used with the lytic enzymes for the quantitative and qualitative determination for the presence of bacteria.

108. Use of stabilizing agents in culture media for growing acid producing bacteria. Kegel, M. A., Wallace, D. L. (1989). Patent number 4,806,479. Disclosed is a bulk starter medium for the propagation of a mother culture of an acid producing bacteria which medium contains a carbohydrate source, a nitrogen containing growth stimulant, a phage control agent and an essentially insoluble or temporarily insolubilized neutralizing agent to which has been added a foodgrade hydrocolloid stabilizing agent.

109. Vaginal suppository for treating group B Streptococcus infection. Fischetti, V., Loomis, L. (2002). Patent number 6,428,784. A composition for treatment of bacterial infections of the vagina is disclosed which comprises a lytic enzyme composition specific for the infecting bacteria, and a carrier for delivering said lytic enzyme. The lytic enzyme is which is specific for the group B Streptococcus, is genectically coded for by a specific bacteriophage.

110. Viral products. Gasson, M. J. (2000). Patent number 6,083,684 (see also 5,763,251). Bacteriophages of food-contaminating or pathogenic bacteria or the lysins thereof are used to kill such bacteria. Examples include lysins from bacteriophages of Listeria monocytogenes and Clostridium tyrobutyricum. Tests for bacterial contamination can be made specific for specific bacteria by using the appropriate bacteriophage or lysin thereof and determining whether cells are lysed thereby.

111. Water system virus detection. Fletcher, J. C., Fraser, A. S., Wells, A. F., Tenoso, H. J. (1978). Patent number 4,118,315. The performance of a waste-water reclamation system is monitored by introducing a non-pathogenic marker virus, bacteriophage F₂, into the waste-water prior to treatment and, thereafter, testing the reclaimed water for the presence of the marker virus. A test sample is first concentrated by absorbing any marker virus onto a cellulose acetate filter in the presence of a trivalent cation at low pH and then flushing the filter with a limited quantity of a glycine buffer solution to desorb any marker virus present on the filter. Photo-optical detection of indirect passive immune agglutination by polystyrene beads indicates the performance of the water reclamation system in removing the marker virus. A closed system provides for concentrating any marker virus, initiating and monitoring the passive immune agglutination reaction, and then flushing the system to prepare for another sample. Peristaltic pumps are provided for volumetric control and for positive fluid displacement. Solenoid valves direct the output from the pumps in preselected routes to accomplish the process for concentrating and detecting the marker virus.

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