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Bacteriophage storage

Victor P Kutyshenko, Galina V Mikoulinskaia, Sergei V Chernyshov, Alexander Y Yegorov, Dmitry A Prokhorov, Vladimir N Uversky
In this work, we studied the effect of the C-terminally attached poly-histidine tag (His-tag), as well as the peculiarities of the protein purification procedure by the immobilized metal affinity chromatography (IMAC) on the activity and structure of the metalloenzyme, l-alanyl-d-glutamate peptidase of bacteriophage T5 (EndoT5), whose zinc binding site and catalytic aspartate are located near the C-terminus. By itself, His-tag did not have a significant effect on either activity or folding of the polypeptide chain, nor on the binding of zinc and calcium ions to the protein...
November 27, 2018: International Journal of Biological Macromolecules
Licheng Huang, Xin Luo, Jingwen Gao, Karl R Matthews
Foodborne illnesses caused by norovirus contaminated fresh produce remain a food safety concern worldwide. In the present study, the impacts of commercial and home processing conditions of strawberries were evaluated for inactivation of the MS2 bacteriophage. MS2 was used as a surrogate of norovirus and was spot inoculated onto strawberries to achieve 6.6 log PFU/g. The inoculated strawberries were washed with tap water, electrolyzed water, or 50 ppm chlorine for 90 s prior to and after storage. After initial washing, the strawberries were separately stored at -20 °C and -80 °C for 30 days...
November 8, 2018: International Journal of Food Microbiology
Alexander B Carstens, Amaru M Djurhuus, Witold Kot, Deborah Jacobs-Sera, Graham F Hatfull, Lars H Hansen
Modern agriculture is expected to face an increasing global demand for food while also needing to comply with higher sustainability standards. Therefore, control of crop pathogens requires new, green alternatives to current methods. Potatoes are susceptible to several bacterial diseases, with infections by soft rot Enterobacteriaceae (SRE) being a significant contributor to the major annual losses. As there are currently no efficient ways of combating SRE, we sought to develop an approach that could easily be incorporated into the potato production pipeline...
November 10, 2018: Viruses
Eva González-Menéndez, Lucía Fernández, Diana Gutiérrez, Ana Rodríguez, Beatriz Martínez, Pilar García
Bacteriophages have been proven as effective antimicrobial agents in the treatment of infectious diseases and in other biocontrol applications including food preservation and disinfection. The extensive use of bacteriophages requires improved methodologies for medium- and long-term storage as well as for easy shipping. To this aim, we have determined the stability of four Staphylococcus phages (phiIPLA88, phiIPLA35, phiIPLA-RODI and phiIPLA-C1C) with antimicrobial potential at different temperatures (20°C/25°C, 4°C, -20°C, -80°C, -196°C) and during lyophilization (freeze drying) using several stabilizing additives (disaccharides, glycerol, sorbitol and skim milk)...
2018: PloS One
Hoang Minh Son, Hoang Minh Duc, Yoshimitsu Masuda, Ken-Ichi Honjoh, Takahisa Miyamoto
Shiga toxin-producing Escherichia coli (STEC) O157:H7 and extended-spectrum beta-lactamase (ESBL) producing E. coli (ESBLEC) are important bacteria of public health concern and frequently isolated from raw beef products. Bacteriophage-based methods have been increasingly exploited to control bacterial contamination in meats. Here, we describe the isolation, characterization, and application of a lytic phage PE37 for the simultaneous bio-control of STEC O157:H7 and ESBLEC. Phage PE37, isolated from the bovine intestine, was morphologically characterized as a member of the Myoviridae family, with a broad host range and great stability under various stress conditions...
September 29, 2018: Applied Microbiology and Biotechnology
Joshua M A Stough, Max Kolton, Joel E Kostka, David J Weston, Dale A Pelletier, Steven W Wilhelm
Sphagnum -dominated peatlands play an important role in global carbon storage and represent significant sources of economic and ecological value. While recent efforts to describe microbial diversity and metabolic potential of the Sphagnum microbiome have demonstrated the importance of its microbial community, little is known about the viral constituents. We used metatranscriptomics to describe the diversity and activity of viruses infecting microbes within the Sphagnum peat bog. The vegetative portions of 6 Sphagnum plants were obtained from a peatland in northern Minnesota and total RNA extracted and sequenced...
September 14, 2018: Applied and Environmental Microbiology
Eva González-Menéndez, Lucía Fernández, Diana Gutiérrez, Daniel Pando, Beatriz Martínez, Ana Rodríguez, Pilar García
The antimicrobial properties of bacteriophages make them suitable food biopreservatives. However, such applications require the development of strategies that ensure stability of the phage particles during food processing. In this study, we assess the protective effect of encapsulation of the Staphylococcus aureus bacteriophage phiIPLA-RODI in three kinds of nanovesicles (niosomes, liposomes, and transfersomes). All these systems allowed the successful encapsulation of phage phiIPLA-RODI with an efficiency ranged between 62% and 98%, regardless of the concentration of components (like phospholipids and surfactants) used for vesicle formation...
September 13, 2018: Viruses
Katarina Gašić, Nemanja Kuzmanović, Milan Ivanović, Anđelka Prokić, Milan Šević, Aleksa Obradović
Xanthomonas euvesicatoria phage KΦ1, a member of Myoviridae family, was isolated from the rhizosphere of pepper plants showing symptoms of bacterial spot. The phage strain expressed antibacterial activity to all X. euvesicatoria strains tested and did not lyse other Xanthomonas spp., nor other less related bacterial species. The genome of KΦ1 is double-stranded DNA of 46.077 bp including 66 open reading frames and an average GC content of 62.9%, representing the first complete genome sequence published for a phage infecting xanthomonads associated with pepper or tomato...
2018: Frontiers in Microbiology
Norton Komora, Carolina Bruschi, Vânia Ferreira, Cláudia Maciel, Teresa R S Brandão, Rui Fernandes, Jorge A Saraiva, Sónia Marília Castro, Paula Teixeira
The application of lytic phages as biocontrol agents is emerging as a promising strategy towards elimination or reduction of foodborne pathogens in a variety of food products. This technology is particularly advantageous for minimally processed and ready-to-eat (RTE) foods. In this study, the potential use of Listex™ P100 combined with high hydrostatic pressure (HPP), to enhance the control of Listeria monocytogenes in food was evaluated. For that, the effect of three pressures (200, 300 or 400 MPa; 5 min, 10 °C) on phage P100 stability was tested when inoculated in six different matrices: phosphate buffered saline (PBS, pH 7...
December 2018: Food Microbiology
Lynn El Haddad, Marie-Josée Lemay, Georges E Khalil, Sylvain Moineau, Claude P Champagne
In an effort to reduce food safety risks, virulent phages are investigated as antibacterial agents for the control of foodborne pathogens. The aim of this study was to evaluate microencapsulation (ME) as a tool to concentrate and store staphylococcal bacteriophages. As a proof of concept, phage Team1 belonging to the Myoviridae family was microencapsulated in alginate gel particles of 0.5 mm (micro-beads) and 2 mm (macro-beads) of diameter. Gel contraction occurred during the hardening period in the CaCl2 solution, and the diameters of the initial alginate droplets shrunk by 16% (micro-beads) and 44% (micro-beads)...
December 2018: Food Microbiology
Alexandros Ch Stratakos, Irene R Grant
Antimicrobial effects of multiple physical, biological and natural interventions on pathogenic Escherichia coli in raw beef were assessed. A cocktail of E. coli strains was inoculated onto gamma-irradiated beef and enumerated immediately after each intervention and during storage at 4 °C for 7 days. Of the physical interventions, silver-containing antimicrobial packaging and ozone gas treatment did not show significant antimicrobial effects, however cold plasma treatment reduced E. coli levels by 0.9 and 1...
December 2018: Food Microbiology
Natasha Bonilla, Jeremy J Barr
A major limitation with traditional phage preparations is the variability in titer, salts, and bacterial contaminants between successive propagations. Here, we introduce the Phage On Tap (PoT) protocol for the quick and efficient preparation of homogenous bacteriophage (phage) stocks. This method produces homogenous, laboratory-scale, high titer (up to 1010-12 PFU/mL), endotoxin reduced phage banks that can be used to eliminate the variability between phage propagations, improve the molecular characterizations of phage, and may be applicable for therapeutic applications...
2018: Methods in Molecular Biology
Jenna Senecal, Annika Nordin, Prithvi Simha, Björn Vinnerås
Over four billion people are discharging untreated human excreta into the environment without any prior treatment, causing eutrophication and spreading disease. The most nutrient rich fraction is the urine. Urine can be collected separately and dehydrated in an alkaline bed producing a nutrient rich fertiliser. However, faecal cross-contamination during the collection risks to introduce pathogens to the urine. The objective of this hygiene assessment was to study the inactivation of five microorganisms (Ascaris suum, Enterococcus faecalis, bacteriophages MS2 and ΦX 174 and Salmonella spp) in alkaline dehydrated urine...
November 1, 2018: Water Research
Naiana Gabiatti, Pingfeng Yu, Jacques Mathieu, Grant W Lu, Xifan Wang, Hangjun Zhang, Hugo M Soares, Pedro J J Alvarez
Bacterial endospores can serve as phage genome protection shells against various environmental stresses to enhance microbial control applications. The genomes of polyvalent lytic Bacillus phages PBSC1 and PBSC2, which infect both B. subtilis subsp. subtilis and B. cereus NRS 248, were incorporated into B. subtilis endospores (without integration into the host chromosome). When PBSC1 and PBSC2 were released from germinating endospores, they significantly inhibited the growth of the targeted opportunistic pathogen B...
September 15, 2018: Applied and Environmental Microbiology
Dongqin Xu, Yanhua Jiang, Lianzhu Wang, Lin Yao, Fengling Li, Yuxiu Zhai, Yuan Zhang
The aim of this study was to evaluate the effectiveness of bacteriophage (phage) SLMP1 to reduce Salmonella Typhimurium on contaminated raw salmon fillets and scallop adductors as a function of Salmonella inoculum level, phage dose, storage temperature, and storage time. Samples were inoculated with 102 and 104 CFU/g Salmonella and then treated with different concentrations of phage SLMP1, followed by incubation at 4, 15, and 25°C, respectively. The results showed that 108 PFU/g was the optimal concentration of phage for the control of Salmonella, which was applied in the following storage experiments over a 7-day period at 4°C, a 4-day period at 15°C, and a 2-day period at 25°C...
August 2018: Journal of Food Protection
Chikage Tanaka, Kohsuke Yamada, Honami Takeuchi, Yoshio Inokuchi, Akiko Kashiwagi, Takahiro Toba
The control of bacterial growth during milk processing is crucial for the quality maintenance of commercial milk and milk products. During a period of cold storage prior to heat treatments, some psychrotrophic bacteria grow and produce extracellular heat-resistant lipases and proteases that cause product defects. The use of lytic bacteriophages (phages) that infect and kill bacteria could be a useful tool for suppressing bacterial growth during this cold storage phase. In this study, we isolated a Pseudomonas lactis strain and a phage from raw cow's milk...
September 15, 2018: Applied and Environmental Microbiology
Li Guo, Ruobing Xu, Lu Gou, Zhichao Liu, Yiming Zhao, Dingxin Liu, Lei Zhang, Hailan Chen, Michael G Kong
Viruses cause serious pathogenic contamination that severely affects the environment and human health. Cold atmospheric-pressure plasma efficiently inactivates pathogenic bacteria; however, the mechanism of virus inactivation by plasma is not fully understood. In this study, surface plasma in argon mixed with 1% air and plasma-activated water was used to treat water containing bacteriophages. Both agents efficiently inactivated bacteriophages T4, Φ174, and MS2 in a time-dependent manner. Prolonged storage had marginal effects on the antiviral activity of plasma-activated water...
September 1, 2018: Applied and Environmental Microbiology
Erica L Vonasek, Angela H Choi, Juan Sanchez, Nitin Nitin
There is a significant unmet need to develop antimicrobial solutions to reduce the risk of contamination in fresh produce. Bacteriophages have been proposed as a potential approach for controlling foodborne pathogens. This study evaluated the combination of edible dip coatings with T7 bacteriophages on whole and cut produce. The evaluation includes an assessment of phage loading, phage storage stability, antimicrobial activity, and phage stability during simulated gastric digestion on sliced cucumbers, sliced apples, and whole cherry tomatoes...
July 2018: Journal of Food Science
Namfon Booncharoen, Skorn Mongkolsuk, Kwanrawee Sirikanchana
Enterococcus faecalis bacteria have been recently reported for their ability to host bacteriophages that are specifically from human sewage, suggesting their application to track human fecal contamination in water resources. However, little is known about the survivability of sewage-specific enterococcal bacteriophages in various water matrices under ambient and storage conditions. In this study, bacteriophages that were derived from the Thailand-isolated E. faecalis strains AIM06 and SR14 exhibited morphologies consistent with the Siphoviridae, Podoviridae, and Myoviridae families...
July 2018: Applied Microbiology and Biotechnology
Jessica Chopyk, Sarah Allard, Daniel J Nasko, Anthony Bui, Emmanuel F Mongodin, Amy R Sapkota
Agricultural ponds have a great potential as a means of capture and storage of water for irrigation. However, pond topography (small size, shallow depth) leaves them susceptible to environmental, agricultural, and anthropogenic exposures that may influence microbial dynamics. Therefore, the aim of this project was to characterize the bacterial and viral communities of pond water in the Mid-Atlantic United States with a focus on the late season (October-December), where decreasing temperature and nutrient levels can affect the composition of microbial communities...
2018: Frontiers in Microbiology
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