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Bacteriophages
Selected AbstractsATTEMPTS TO UTILIZE BACTERIOPHAGE TO COMBAT SALMONELLA ENTERICA SEROVAR ENTEMTIDIS INFECTION IN CHICKENSJOURNAL OF FOOD SAFETY, Issue 1 2001IAN B. SKLAR ABSTRACT Bacteriophage capable of lysing a nalidixic acid-resistant Salmonella enterica serovar Enteritidis strain (SeE Nalr) were tested for the ability to reduce cecal Salmonella counts in young chickens infected with the bacterium. Qualitative analysis of cloacal swabs suggested that phage treatment can possibly reduce shedding of SeE Nalr, but average SeE Nalr counts of between 105 and 107 cfu of SeE Nalr per g of cecum were obtained even from phage-treated 14-day old birds and even when more than 107 plaque forming units of phage were present per gram of cecal content. The average cecal SeE Nalr counts were generally between 0.3 and 1.3 orders of magnitude lower in phage-treated chickens than in untreated controls birds. The difference in counts was statistically not significant in three animal trials, but significant in two trials using feed particles as delivery vehicles for the phage. Although some of the SeE Nalr in the cecae of phage-treated chickens had developed resistance to some of the phage used, factors other than phage resistance must have contributed to the failure of the phage to substantially reduce SeE Nalr counts. [source] EFFECT OF HIGH PRESSURE ON LACTOCOCCAL BACTERIOPHAGESJOURNAL OF FOOD SAFETY, Issue 1 2009M. DILEK ABSTRACT Four different host-specific lactococcal bacteriophages were subjected to high hydrostatic pressure and heat treatments. Pressure treatments were done at room temperature at 300 and 350 MPa for 5,40 min. Complete inactivation of bacteriophages was observed starting at 350 MPa for 20-min treatment at room temperature. The effect of heat on the bacteriophages was analyzed by heat treatment at 71.7C for predetermined lengths of time (1,5 min). Decrease in bacteriophage number was observed after 3 min of heat treatment at 71.7C. Pressure treatment at 350 MPa/5 min and heat treatment at 71.7C/3 min were both found to be effective for the inactivation of lactococcal bacteriophages. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis analysis indicated that protein profiles of pressure-treated (350 MPa, 25 min) bacteriophages were altered. PRACTICAL APPLICATIONS Bacteriophages are still a problem for the production of fermented dairy products, as there has not been a process to eliminate them completely from the fermentation environment. Processes such as pasteurization are not adequate to eliminate bacteriophages. However, new food preservation methods have been developed, one of which is high hydrostatic pressure (HHP) processing. HHP has potential application for the inactivation of viruses. Here, we demonstrate the application of HHP to inactivate the bacteriophages of dairy starter culture Lactococcus in comparison with heat treatment. [source] Translation repression by an RNA polymerase elongation complexMOLECULAR MICROBIOLOGY, Issue 3 2004Helen R. Wilson Summary Bacteriophage , N and bacterial Nus proteins together with a unique site NUT in the leader of the early viral N gene transcript bind RNA polymerase (RNAP) and form a highly processive antitermination complex; N bound at NUT also represses N translation. In this study, we investigate whether N and NUT cause N translation repression as part of the antitermination complex by testing conditions that inhibit the formation of the N-modified transcription complex for their effect on N-mediated translation repression. We show that nus and nut mutations that in combination destabilize multiple interactions in the antitermination complex prevent N-mediated translation repression. Likewise, transcription of the nut-N region by T7 RNAP, which does not lead to the assembly of an effective antitermination complex when N is supplied, eliminates translation repression. We also demonstrate that a unique mutant , subunit of RNAP reduces N-mediated translation repression, and that overexpression of transcription factor NusA suppresses this defect. We conclude that the N-modified RNAP transcription complex is necessary to repress N translation. [source] Enhanced matrix-assisted laser desorption/ionization time-of-flight mass spectrometric analysis of bacteriophage major capsid proteins with , -mercaptoethanol pretreatmentRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 1 2010Casey R. McAlpin Bacteriophage (phage) proteins have been analyzed previously with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). However, analysis of phage major capsid proteins (MCPs) has been limited by the ability to reproducibly generate ions from MCP monomers. While the acidic conditions of MALDI-TOF MS sample preparation have been shown to aid in disassembly of some phage capsids, many require further treatment to successfully liberate MCP monomers. The findings presented here suggest that , -mercaptoethanol reduction of the disulfide bonds linking phage MCPs prior to mass spectrometric analysis results in significantly increased MALDI-TOF MS sensitivity and reproducibility of Yersinia pestis -specific phage protein profiles. Copyright © 2009 John Wiley & Sons, Ltd. [source] Crystallization and preliminary crystallographic analysis of poly-,-glutamate hydrolase from bacteriophage ,NIT1ACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 9 2009Zui Fujimoto Particular Bacillus subtilis strains produce a capsular polypeptide poly-,-glutamate (,-PGA) that functions as a physical barrier against bacteriophage infection. Bacteriophage ,NIT1 can infect B. subtilis and produces a novel ,-PGA hydrolase PghP. PghP was overexpressed, purified and crystallized by the sitting-drop vapour-diffusion method. The crystals diffracted to a resolution of 2.4,Å using a synchrotron X-ray source and were found to belong to space group P3121 or P3221. [source] A Recombinant Bacteriophage-Based Assay for the Discriminative Detection of Culturable and Viable but Nonculturable Escherichia coli O157:H7BIOTECHNOLOGY PROGRESS, Issue 3 2006Raheela Awais A previously green fluorescent protein (GFP)-labeled PP01 virulent bacteriophage, specific to Escherichia coli O157:H7, was used to construct lysozyme-inactivated GFP-labeled PP01 phage (PP01e - /GFP). The new recombinant phage lacked lytic activity because of the inactivation of gene e, which produces the lysozyme responsible for cell lysis. Gene e was inactivated by inserting an amber stop codon. Prolonged incubation ofE. coli O157:H7 cells with PP01e - /GFP did not lead to cell lysis, while the propagation of PP01e - /GFP in host cells increased the intensity of green fluorescence. Retention of cell morphology and increase in fluorescence enabled the direct visualization and enumeration of E. coli O157:H7 cells within an hour. The PP01e - /GFP system, when combined with nutrient uptake analysis, further allowed the discriminative detection of culturable, viable but nonculturable (VBNC), and dead cells in the stress-induced aquatic environment. Stress-induced cells, which retained culturability, allowed phage propagation and produced bright green florescence. Nonculturable cells (VBNC and dead) allowed only phage adsorption but no proliferation and remained low fluorescent. The low-fluorescent nonculturable cells were further differentiated into VBNC and dead cells on the basis of nutrient uptake analysis. The low-fluorescent cells, which grew in size by nutrient incorporation during prolonged incubation in nutrient medium, were defined as metabolically active and in the VBNC state. The elongated VBNC cells were then easily recognizable from dead cells. The proposed assay enabled the detection and quantification of VBNC cells. Additionally, it revealed the proportion of culturable to VBNC cells within the population, as opposed to conventional techniques, which demonstrate VBNC cells as a differential value of the total viable count and the culturable cell count. [source] A Coming of 'Phage: Bacteriophages in BiotechnologyJOURNAL OF CHEMICAL TECHNOLOGY & BIOTECHNOLOGY, Issue 7 2001Richard J Sharp Copyright © 2001 Society of Chemical Industry [source] Phage-mediated transfer of virulence genesJOURNAL OF CHEMICAL TECHNOLOGY & BIOTECHNOLOGY, Issue 7 2001Jon R Saunders Abstract Bacteriophages as accessory genetic elements play a crucial role in the dissemination of genes and the promotion of genetic diversity within bacterial populations. Such horizontal transfer of DNA is critical in the emergence of new pathogenic organisms, through the dissemination of genes encoding virulence factors such as toxins, adhesins and agressins. Phages can transfer genes that are not necessary for bacteriophage persistence and are generally recognised by their ability to convert their host bacteria to new phenotypes. This phenomenon is known as phage conversion. If such converting genes encode for virulence factors, the consequences of phage infection may include increased virulence of the host bacteria, and the conversion of a non-pathogenic strain to a potentially dangerous pathogen. A number of virulence factors in bacteria causing diseases in plants, animals and humans are encoded by converting phages, the vast majority of which are temperate as opposed to lytic in nature. © 2001 Society of Chemical Industry [source] Bacteriophages: biology and historyJOURNAL OF CHEMICAL TECHNOLOGY & BIOTECHNOLOGY, Issue 7 2001Richard Sharp Abstract Bacteriophages were initially considered to offer the key to the control of bacterial infections; early studies, however, proved largely unsuccessful. In the 1940s and 1950s, pioneering studies into the structure and physiology of host/phage interactions laid the basis for the development of molecular biology and a spectrum of new biotechnologically-based industries. Bacteriophages able to infect most procaryotic groups of organisms have been isolated, and are readily isolated from soil, water, and sewage and most environments colonised by bacteria. Ecologically, phages are as varied and as versatile as their hosts with some able to survive extremes of temperature (up to 95,°C) and extremes of pH as low as pH 1. © 2001 Society of Chemical Industry [source] EFFECT OF HIGH PRESSURE ON LACTOCOCCAL BACTERIOPHAGESJOURNAL OF FOOD SAFETY, Issue 1 2009M. DILEK ABSTRACT Four different host-specific lactococcal bacteriophages were subjected to high hydrostatic pressure and heat treatments. Pressure treatments were done at room temperature at 300 and 350 MPa for 5,40 min. Complete inactivation of bacteriophages was observed starting at 350 MPa for 20-min treatment at room temperature. The effect of heat on the bacteriophages was analyzed by heat treatment at 71.7C for predetermined lengths of time (1,5 min). Decrease in bacteriophage number was observed after 3 min of heat treatment at 71.7C. Pressure treatment at 350 MPa/5 min and heat treatment at 71.7C/3 min were both found to be effective for the inactivation of lactococcal bacteriophages. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis analysis indicated that protein profiles of pressure-treated (350 MPa, 25 min) bacteriophages were altered. PRACTICAL APPLICATIONS Bacteriophages are still a problem for the production of fermented dairy products, as there has not been a process to eliminate them completely from the fermentation environment. Processes such as pasteurization are not adequate to eliminate bacteriophages. However, new food preservation methods have been developed, one of which is high hydrostatic pressure (HHP) processing. HHP has potential application for the inactivation of viruses. Here, we demonstrate the application of HHP to inactivate the bacteriophages of dairy starter culture Lactococcus in comparison with heat treatment. [source] Use of Bacteriophages to Control Salmonella in Experimentally Contaminated Sprout SeedsJOURNAL OF FOOD SCIENCE, Issue 5 2004S. Pao ABSTRACT: Trials were conducted to evaluate the potential for using bacteriophages to control Salmonella in sprouting seeds. Two phages (Phage-A, capable of lysing S. Typhimurium and S. Enteritidis, and Phage-B, capable of lysing S. Montevideo) were isolated and characterized as members of the Myoviridae and Siphoviridae families, respectively. Salmonella counts increased in all inoculated seeds during soaking and mustard seeds supported greater growth of the inoculated Salmonella than broccoli seeds. A 1.37 log suppression of Salmonella growth was achieved by applying Phage-A on mustard seeds. The mixture of Phage-A and Phage-B caused a 1.50 log suppression of Salmonella growth in the soaking water of broccoli seeds. Host specificity observed in the study stresses the importance of developing phage mixtures that can control a broad range of potential contaminants. [source] Prevalence of Salmonella infecting bacteriophages associated with Ontario pig farms and the holding area of a high capacity pork processing facilityJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 13 2010Sunan Wang Abstract BACKGROUND: There is interest in applying bacteriophages to control Salmonella in pig production and pork processing. The following reports on the prevalence of Salmonella infecting bacteriophages within Ontario pig farms and associated with the holding area of a pork slaughterhouse. RESULTS:Salmonella infecting bacteriophages were present in 30 and 28 of the effluent manure samples collected from 36 farms using S. Typhimurium DT104 or S. Heidelberg as host cell respectively. Bacteriophages were recovered in 95,100% of the 48 samples taken from holding pens within a high capacity slaughterhouse over a 12 month period. Bacteriophages isolated from farms exhibited similar host ranges which differed to that of slaughterhouse isolates. Salmonella (n = 21) from the slaughterhouse were susceptible to the endogenous bacteriophages. Despite being susceptible to the resident phages, the Salmonella populations were found to be genetically stable with the same genotypes being recovered over successive visits. Salmonella isolated from the farms were frequently resistant to the endogenous phages. CONCLUSIONS: Bacteriophages are prevalent in the pig slaughterhouse environment although they do not have a significant impact on the genetic structure of Salmonella populations. However, there was evidence that the Salmonella population structure on farms is influenced by the presence of infecting phages. Copyright © 2010 Society of Chemical Industry [source] Isolation and characterization of bacteriophages specific for Campylobacter jejuniMICROBIOLOGY AND IMMUNOLOGY, Issue 10 2009Sunyoung Hwang ABSTRACT Human infection by Campylobacter jejuni is mainly through the consumption of contaminated poultry products, which results in gastroenteritis and, rarely, bacteremia and polyneuropathies. In this study, six C. jejuni -specific bacteriophages (CPS1,6) were isolated by the spot-on-the-lawn technique from chicken samples in Korea and characterized for potential use as biocontrol agents. All isolated bacteriophages exhibited a high specificity, being able to lyse only C. jejuni, but not other Gram,negative bacteria, including C. coli, Escherichia coli, Salmonella spp., and Gram,positive bacteria. Bacteriophages contain an icosahedral head and a contractile tail sheath in transmission electron microscopy, and possess ds-DNA with an average genome size of approximately 145 kb; therefore, all bacteriophages are categorized into the Myoviridae family. Bacterial lysis studies in liquid media revealed that CPS2 could be used to control the growth of C. jejuni. [source] Bacteriophage WO-B and Wolbachia in natural mosquito hosts: infection incidence, transmission mode and relative densityMOLECULAR ECOLOGY, Issue 9 2006N. CHAUVATCHARIN Abstract Bacteriophages of Wolbachia bacteria have been proposed as a potential transformation tool for genetically modifying mosquito vectors. In this study, we report the presence of the WO-B class of Wolbachia -associated phages among natural populations of several mosquito hosts. Eighty-eight percent (22/25) of Wolbachia -infected mosquito species surveyed were found to contain WO-B phages. WO-B phage orf7 sequence analysis suggested that a single strain of WO-B phage was found in most singly (23/24) or doubly (1/1) Wolbachia -infected mosquitoes. However, the single Wolbachia strain infecting Aedes perplexus was found to harbour at least two different WO-B phages. Phylogenetic analysis suggested that horizontal transmission of WO-B phages has occurred on an evolutionary scale between the Wolbachia residing in mosquitoes. On an ecological scale, a low trend of co-transmission occurred among specific WO-B phages within Wolbachia of each mosquito species. Assessment of the density of WO-B phage by real-time quantitative polymerase chain reaction (RTQ-PCR) revealed an average relative density of 7.76 × 105± 1.61 × 105 orf7 copies per individual mosquito for a single Wolbachia strain infecting mosquitoes, but a threefold higher density in the doubly Wolbachia-infected Aedes albopictus. However, the average combined density of WO-B phage(s) did not correlate with that of their Wolbachia hosts, which varied in different mosquito species. We also confirmed the presence of WO-B-like virus particles in the laboratory colony of Ae. albopictus (KLPP) morphologically, by transmission electron microscopy (TEM). The viral-like particles were detected after purification and filtration of Ae. albopictus ovary extract, suggesting that at least one WO-B-like phage is active (temperate) within the Wolbachia of this mosquito vector. Nevertheless, the idea of utilizing these bacteriophages as transformation vectors still needs more investigation and is likely to be unfeasible. [source] Extensive phage dynamics in Staphylococcus aureus contributes to adaptation to the human host during infectionMOLECULAR MICROBIOLOGY, Issue 6 2006Christiane Goerke Summary Bacteriophages serve as a driving force in microbial evolution, adaptation to new environments and the pathogenesis of human bacterial infections. In Staphylococcus aureus phages encoding immune evasion molecules (SAK, SCIN, CHIPS), which integrate specifically into the ,-haemolysin (Hlb) gene, are widely distributed. When comparing S. aureus strain collections from infectious and colonizing situations we could detect a translocation of sak -encoding phages to atypical genomic integration sites in the bacterium only in the disease-related isolates. Additionally, significantly more Hlb producing strains were detected in the infectious strain collection. Extensive phage dynamics (intragenomic translocation, duplication, transfer between hosts, recombination events) during infection was shown by analysing cocolonizing and consecutive isolates of patients. This activity leads to the splitting of the strain population into various subfractions exhibiting different virulence potentials (Hlb-production and/or production of immune evasion molecules). Thus, phage-inducing conditions and strong selection for survival of the bacterial host after phage movement are typical for the infectious situation. Further in vitro characterization of phages revealed that: (i) SAK is encoded not only on serogroup F phages showing a conserved tropism for hlb but also on serogroup B phages which always integrate in a distinct intergenic region, (ii) the level of sak transcription correlates to phage inducibility but is independent of the phage localization in the chromosome, and (iii) phages can be stabilized extra-chromosomally during their life cycle. [source] Bacteriophages induced from lysogenic root canal isolates of Enterococcus faecalisMOLECULAR ORAL MICROBIOLOGY, Issue 4 2009R. H. Stevens Introduction:, Bacterial viruses play crucial roles in the pathogenesis of many systemic diseases. They are known to inhabit the oral cavity, both as free virions and as prophages in lysogenic bacterial strains; however, there has been no report of bacteriophages in endodontic infections. In this study, we sought to detect, isolate, and describe temperate bacteriophages harbored by Enterococcus faecalis strains isolated from endodontic infections. Methods: Ten E. faecalis strains were isolated from root canals of teeth undergoing retreatment following unsuccessful endodontic therapy. Mitomycin C was used to induce any prophages present in the bacterial isolates. The induced phages were purified and examined using electron microscopy. The DNA extracted from one of the phage isolates was subjected to restriction endonuclease digestion and agarose electrophoresis analysis. Results:, Lysogeny was demonstrated in 4 of the 10 E. faecalis strains. Three of the lysogenic strains yielded phages exhibiting a Siphoviridae morphology, with long, non-contractile tails 130 nm in length, and spherical/icosahedral heads 41 nm in diameter. The virus induced from the fourth lysogenic E. faecalis strain had a contractile tail characteristic of Myoviridae. Restriction endonuclease analysis of NsiI and NdeI DNA fragments from one of the Siphoviridae phage isolates (phage ,Ef11) indicated a genome size of approximately 41 kbp. Conclusion:, This is the first report of lysogenic bacteria and their inducible viruses in infected root canals. [source] Characterization of a Vibrio cholerae phage isolated from the coastal water of PeruENVIRONMENTAL MICROBIOLOGY, Issue 5 2003Miguel Talledo Summary A Vibrio cholerae bacteriophage, family Myoviridae, was isolated from seawater collected from the coastal water of Lima, Peru. Genome size was estimated to be 29 kbp. The temperate phage was specific to V. cholerae and infected 12/13 V. cholerae O1 strains and half of the four non-O1/non-O139 strains tested in this study. Vibrio cholerae O139 strains were resistant to infection and highest infection rates were obtained in low nutrient media amended with NaCl or prepared using seawater as diluent. [source] COEVOLUTION DRIVES TEMPORAL CHANGES IN FITNESS AND DIVERSITY ACROSS ENVIRONMENTS IN A BACTERIA,BACTERIOPHAGE INTERACTIONEVOLUTION, Issue 8 2008Samantha E. Forde Coevolutionary interactions are thought to play a crucial role in diversification of hosts and parasitoids. Furthermore, resource availability has been shown to be a fundamental driver of species diversity. Yet, we still do not have a clear understanding of how resource availability mediates the diversity generated by coevolution between hosts and parasitoids over time. We used experiments with bacteria and bacteriophage to test how resources affect variation in the competitive ability of resistant hosts and temporal patterns of diversity in the host and parasitoid as a result of antagonistic coevolution. Bacteria and bacteriophage coevolved for over 150 bacterial generations under high and low-resource conditions. We measured relative competitive ability of the resistant hosts and phenotypic diversity of hosts and parasitoids after the initial invasion of resistant mutants and again at the end of the experiment. Variation in relative competitive ability of the hosts was both time- and environment-dependent. The diversity of resistant hosts, and the abundance of host-range mutants attacking these phenotypes, differed among environments and changed over time, but the direction of these changes differed between the host and parasitoid. Our results demonstrate that patterns of fitness and diversity resulting from coevolutionary interactions can be highly dynamic. [source] Developing live Shigella vaccines using , Red recombineeringFEMS IMMUNOLOGY & MEDICAL MICROBIOLOGY, Issue 3 2006Ryan T. Ranallo Abstract Live attenuated Shigella vaccines have shown promise in inducing protective immune responses in human clinical trials and as carriers of heterologous antigens from other mucosal pathogens. In the past, construction of Shigella vaccine strains relied on classical allelic exchange systems to genetically engineer the bacterial genome. These systems require extensive in vitro engineering of long homologous sequences to create recombinant replication-defective plasmids or phage. Alternatively, the ,red recombination system from bacteriophage facilitates recombination with as little as 40 bp of homologous DNA. The process, referred to as recombineering, typically uses an inducible ,red operon on a temperature-sensitive plasmid and optimal transformation conditions to integrate linear antibiotic resistance cassettes flanked by homologous sequences into a bacterial genome. Recent advances in recombineering have enabled modification of genomic DNA from bacterial pathogens including Salmonella, Yersinia, enteropathogenic Escherichia coli, or enterohemorrhagic E. coli and Shigella. These advances in recombineering have been used to systematically delete virulence-associated genes from Shigella, creating a number of isogenic strains from multiple Shigella serotypes. These strains have been characterized for attenuation using both in vivo and in vitro assays. Based on this data, prototypic Shigella vaccine strains containing multiple deletions in virulence-associated genes have been generated. [source] The life cycles of the temperate lactococcal bacteriophage ,LC3 monitored by a quantitative PCR methodFEMS MICROBIOLOGY LETTERS, Issue 1 2000Merete Lunde Abstract We present here a new and general approach for monitoring the life cycles of temperate bacteriophages which establish lysogeny by inserting their genomes site-specifically into the bacterial host chromosome. The method is based on quantitative amplification of specific DNA sites involved in various cut-and-join events during the life cycles of the phages (i.e. the cos, attP, attB, attL and attR sites) with the use of sequence-specific primers. By comparing the amounts of these specific DNA sites at different intervals, we were able to follow the development of the lytic and lysogenic life cycles of the temperate lactococcal bacteriophage ,LC3 after infection of its bacterial host Lactococcus lactis ssp. cremoris IMN-C18. [source] DNA bending and looping in the transcriptional control of bacteriophage ,29FEMS MICROBIOLOGY REVIEWS, Issue 5 2010Ana Camacho Abstract Recent studies on the regulation of phage ,29 gene expression reveal new ways to accomplish the processes required for the orderly gene expression in prokaryotic systems. These studies revealed a novel DNA-binding domain in the phage main transcriptional regulator and the nature and dynamics of the multimeric DNA,protein complex responsible for the switch from early to late gene expression. This review describes the features of the regulatory mechanism that leads to the simultaneous activation and repression of transcription, and discusses it in the context of the role of the topological modification of the DNA carried out by two phage-encoded proteins working synergistically with the DNA. [source] Genetically Engineered Phage Fibers and Coatings for Antibacterial ApplicationsADVANCED FUNCTIONAL MATERIALS, Issue 2 2010Joan Y. Mao Abstract Multifunctionality can be imparted to protein-based fibers and coatings via either synthetic or biological approaches. Here, potent antimicrobial functionality of genetically engineered, phage-based fibers and fiber coatings, processed at room temperature, is demonstrated. Facile genetic engineering of the M13 virus (bacteriophage) genome leverages the well-known antibacterial properties of silver ions to kill bacteria. Predominant expression of negatively charged glutamic acid (E3) peptides on the pVIII major coat proteins of M13 bacteriophage enables solution-based, electrostatic binding of silver ions and subsequent reduction to metallic silver along the virus length. Antibacterial fibers of micrometer-scale diameters are constructed from such an E3-modified phage via wet-spinning and glutaraldehyde-crosslinking of the E3-modified viruses. Silverization of the free-standing fibers is confirmed via energy dispersive spectroscopy and inductively coupled plasma atomic emission spectroscopy, showing ,0.61,µg cm,1 of silver on E3,Ag fibers. This degree of silverization is threefold greater than that attainable for the unmodified M13,Ag fibers. Conferred bactericidal functionality is determined via live,dead staining and a modified disk-diffusion (Kirby,Bauer) measure of zone of inhibition (ZoI) against Staphylococcus epidermidis and Escherichia coli bacterial strains. Live,dead staining and ZoI distance measurements indicate increased bactericidal activity in the genetically engineered, silverized phage fibers. Coating of Kevlar fibers with silverized E3 phage exhibits antibacterial effects as well, with relatively smaller ZoIs attributable to the lower degree of silver loading attainable in these coatings. Such antimicrobial functionality is amenable to rapid incorporation within fiber-based textiles to reduce risks of infection, biofilm formation, or odor-based detection, with the potential to exploit the additional electronic and thermal conductivity of fully silverized phage fibers and coatings. [source] Multiple displacement amplification to create a long-lasting source of DNA for genetic studies,HUMAN MUTATION, Issue 7 2006Lovisa Lovmar Abstract In many situations there may not be sufficient DNA collected from patient or population cohorts to meet the requirements of genome-wide analysis of SNPs, genomic copy number polymorphisms, or acquired copy number alternations. When the amount of available DNA for genotype analysis is limited, high performance whole-genome amplification (WGA) represents a new development in genetic analysis. It is especially useful for analysis of DNA extracted from stored histology slides, tissue samples, buccal swabs, or blood stains collected on filter paper. The multiple displacement amplification (MDA) method, which relies on isothermal amplification using the DNA polymerase of the bacteriophage ,29, is a recently developed technique for high performance WGA. This review addresses new trends in the technical performance of MDA and its applications to genetic analyses. The main challenge of WGA methods is to obtain balanced and faithful replication of all chromosomal regions without the loss of or preferential amplification of any genomic loci or allele. In multiple comparisons to other WGA methods, MDA appears to be most reliable for genotyping, with the most favorable call rates, best genomic coverage, and lowest amplification bias. Hum Mutat 27(7), 603,614, 2006. © 2006 Wiley-Liss, Inc. [source] The ability of two different Vibrio spp. bacteriophages to infect Vibrio harveyi, Vibrio cholerae and Vibrio mimicusJOURNAL OF APPLIED MICROBIOLOGY, Issue 4 2004M. Payne Abstract Aims:, To determine the host range of the Vibrio harveyi myovirus-like bacteriophage (VHML) and the cholera toxin conversion bacteriophage (CTX ,) within a range of Vibrio cholerae and V. mimicus and V. harveyi, V. cholerae and V. mimicus isolates respectively. Methods and Results:, Three V. harveyi, eight V. cholerae and five V. mimicus isolates were incubated with VHML and CTX ,. Polymerase chain reaction (PCR) was used to determine the presence of VHML and CTX , in infected isolates. We demonstrated that it was possible to infect one isolate of V. cholerae (isolate ACM #2773/ATCC #14035) with VHML. This isolate successfully incorporated VHML into its genome as evident by positive PCR amplification of the sequence coding part of the tail sheath of VHML. Attempts to infect all other V. cholerae and V. mimicus isolates with VHML were unsuccessful. Attempts to infect V. cholerae non-01, V. harveyi andV. mimicus isolates with CTX , were unsuccessful. Conclusions:, Bacteriophage infection is limited by bacteriophage-exclusion systems operating within bacterial strains and these systems appear to be highly selective. One system may allow the co-existence of one bacteriophage while excluding another. VHML appears to have a narrow host range which may be related to a common receptor protein in such strains. The lack of the vibrio pathogenicity island bacteriophage (VPI ,) in the isolates used in this study may explain why infections with CTX , were unsuccessful. Significance and Impact of the Study:, The current study has demonstrated that Vibrio spp. bacteriophages may infect other Vibrio spp. [source] A new bacteriophage, VHML, isolated from a toxin-producing strain of Vibrio harveyi in tropical AustraliaJOURNAL OF APPLIED MICROBIOLOGY, Issue 4 2000H.J. Oakey Some strains of Vibrio harveyi are known to be pathogenic for fish and many invertebrates including crustaceans. Despite their importance, their modes of virulence have yet to be fully elucidated. Here, we present a previously unreported bacteriophage extracted from a toxin-producing strain of V. harveyi isolated from moribund prawn larvae in tropical Australia. Classification into the family Myoviridae was based upon morphological characteristics (an icosahedral head, a neck/collar region and a sheathed rigid tail) and nucleic acid characteristics (double-stranded linear DNA). We have termed the bacteriophage VHML (Vibrio Harveyi Myovirus Like). VHML is a temperate bacteriophage that has a narrow host range and shows an apparent preference for V. harveyi above other vibrios (63 Vibrio isolates tested) and other genera (10 other genera were tested). The conventional methods for phage concentration and extraction of nucleic acids from phage particles were not efficient and the alternative methods that were used are discussed. [source] Myth and reality: practical test system for the measurement of anti-DNA antibodies in the diagnosis of systemic lupus erythematosus (SLE)JOURNAL OF CLINICAL LABORATORY ANALYSIS, Issue 2 2010Laura J. McCloskey Abstract The myth persists that only the labor intensive Farr radioimmunoassay and Crithidia luciliae immunofluorescence (CL-IFA) are systemic lupus erythematosus (SLE)-specific tests. We compared them to ELISA with bacteriophage , DNA (EL-dsDNA) and denatured calf thymus DNA (EL-ssDNA). By percentile ranking, the specificity cut-off level was set both out of clinical context (SOCC) on 100 blood bank donors, and in clinical context (SICC) on 100 patients with either rheumatoid arthritis or scleroderma (50/50). Clinical sensitivity was calculated on 100 random SLE patients. At 95% SICC, the sensitivity of Farr, CL-IFA, EL-dsDNA, and EL-ssDNA was similar (95%CI): 76% (66,84), 76% (66,84), 63% (53,72), and 75% (65,83), respectively; 87% of the patients were positive by at least one method and 55%by all methods. At 99% SICC, the sensitivity was also similar (95% CI): 57% (47,67), 47% (37,57), 58% (47,67), and 43% (33,53), respectively. The areas under ROC curve were similar (95% CI) when patients were used as controls for specificity. At 99% SOCC, EL-ssDNA identified 89% positive, 2 negative but positive by another method at 95% SICC, and 9 negative (i.e. 89/2/9), followed by CL-IFA (80/6/14), Farr (76/12/12), and EL-dsDNA (64/23/13). Thus, at relatively low cost and easy automation, under the same conditions of specificity, the two ELISA tests combined were at least as good, if not superior, to CL-IFA or Farr: they showed similar clinical sensitivity and also identified more patients with anti-DNA antibodies. J. Clin. Lab. Anal. 24:77,84, 2010. © 2010 Wiley-Liss, Inc. [source] Facile detection of specific RNA-polypeptide interactions by MALDI-TOF mass spectrometryJOURNAL OF PEPTIDE SCIENCE, Issue 8 2008Maki Sugaya Abstract A simple method for the detection of specific RNA-polypeptide interactions using MALDI-TOF mass spectroscopy is described. Instead of direct observation of the RNA-polypeptide complex, we attempted the indirect observation of the binding event by focusing on the disappearance of the free polypeptide signal upon interaction with RNA. As a result, specific binding of the Rev-response element (RRE) RNA of the HIV with two RRE-binding peptide aptamers, DLA and RLA peptides, as well as the bacteriophage , boxB RNA with the , N peptide was observed. We also show that specific RNA-binding peptides can be identified from a mixture of peptides with varying RNA-binding affinity, showing that the method could be applied to high-throughput screening from simple peptide libraries. The method described in this study provides a quick and simple method for detecting specific RNA,polypeptide interactions that avoids difficulties associated with direct observation of RNA and RNA,polypeptide complexes, which may find various applications in the analysis of RNA,polypeptide interactions and in the identification of novel RNA-binding polypeptides. Copyright © 2008 European Peptide Society and John Wiley & Sons, Ltd. [source] Cyclic peptides selected by phage display mimic the natural epitope recognized by a monoclonal anti-colicin A antibodyJOURNAL OF PEPTIDE SCIENCE, Issue 11 2004Stephane Coulon Abstract A 10-mer random peptide library displayed on filamentous bacteriophage was used to determine the molecular basis of the interaction between the monoclonal anti-colicin A antibody 1C11 and its cognate epitope. Previous studies established that the putative epitope recognized by 1C11 antibody is composed of amino acid residues 19,25 (RGSGPEP) of colicin A. Using the phage display technique it was confirmed that the epitope of 1C11 antibody was indeed restricted to residues 19,25 and the consensus motif RXXXPEP was identified. Shorter consensus sequences (RXXPEP, RXXEP, KXXEP) were also selected. It was also demonstrated that the disulfide bond found in one group of the selected peptides was crucial for 1C11 antibody recognition. It was shown that cyclization of the peptides by disulfide bond formation could result in a structure that mimics the natural epitope of colicin A. Copyright © 2004 European Peptide Society and John Wiley & Sons, Ltd. [source] Vibrio harveyi: a significant pathogen of marine vertebrates and invertebratesLETTERS IN APPLIED MICROBIOLOGY, Issue 2 2006B. Austin Abstract Vibrio harveyi, which now includes Vibrio carchariae as a junior synonym, is a serious pathogen of marine fish and invertebrates, particularly penaeid shrimp. In fish, the diseases include vasculitis, gastro-enteritis and eye lesions. With shrimp, the pathogen is associated with luminous vibriosis and Bolitas negricans. Yet, the pathogenicity mechanisms are imprecisely understood, with likely mechanisms involving the ability to attach and form biofilms, quorum sensing, various extracellular products including proteases and haemolysins, lipopolysaccharide, and interaction with bacteriophage and bacteriocin-like substances. [source] Inhibition of bacteriophage K proliferation on Staphylococcus aureus in raw bovine milkLETTERS IN APPLIED MICROBIOLOGY, Issue 3 2005S. O'Flaherty Abstract Aims:, To assess the ability of staphylococcal bacteriophage K to inhibit Staphylococcus aureus in raw milk. Methods and Results:, The ability of bacteriophage (phage) to replicate in milk is important in situations where phage might be used as a therapeutic for bovine mastitis. Phage K was able to replicate normally, leading to elimination of the host culture in milk, which had been previously heat-treated. When raw milk was used under identical conditions, the phages were unable to replicate. Phage adsorption assays were performed and these demonstrated that adsorption of phage was significantly reduced in the raw milk while it was restored in the heat-treated sample (86·50% compared with 99·96% adsorption respectively). When confocal microscopy with a Live/Dead Bac light staining system was employed, it was observed that in raw milk S. aureus formed clusters associated with fat globules, while in heat-treated milk, bacterial agglutination had not occurred. Conclusions:, Raw milk inhibits staphylococcal phage K proliferation. Significance and Impact of the Study:, This observation has implications for the exploitation of staphylococcal therapeutic phage in milk. [source] |