Posters and presentation information
Thank you for considering to present your work as a poster at Phages 2018.
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Posters will be displayed for the full duration of the conference.Titles of accepted poster abstracts will be displayed below.
Accepted posters
(Presenters in Bold)
If your abstract has been accepted for presentation but it does not appear in the list below, please let us know as soon as possible by email at PhageOxford@gmail.com.
A selection of posters can be downloaded on this link (password protected).
A cocktail phage therapy in controlling bacterial disease in broiler feeding compared with antiobiotic therapy
Xinyong Du1, Junwei Liu2, Xianshen Li2
1Agricultural school, Univeristy of Liaocheng, Dongchangfu District, Liaocheng, Shangdong Province, 252000, China PR
2Runda biological engineering institute, Jimo, Qingdao City, Shandong Province, 266200, China PR
A cocktail phage (host bacteria were E. coli and Samonella) therapy compared with antibiotic therapy was applied in broiler farm to control bacterial infectoins in four pheasantries with 15,000 chicken. The phage cocktail therapy compared with antibiotic was carried out at 19~20 days age of chickens when bacterial pathogens were active. Both phages and antibiotics were oral administration via the waterline, and the dosages of phage therapy were: 0.1, 0.3 and 0.5 mL per chicken with an initial phage titer of 1×108 pfu/mL. The amount of death chickens was recorded daily, and during the experiment, those deaths were dissected to find out the symptom of pericarditis, which was regarded as the consequence of bacterial infections. The records were confined 5 days before and after the therapy period. The results showed that: the antibiotic-therapy decreased the average number of daily death chickens from 14 to 11.2 before and after the therapy, while there were significant differences in the phage-therapy groups. Both 0.1 mL per chicken and 0.3 mL per chicken dosage groups successfully decreased the death average numbers from 16 to 10, and 30 to 16, but the 0.5 mL per chicken dosage group failed to decrease the death number, the result of average death chicken numbers were 14.5 to 29. After disscetion of the death chickens, the rate of pericarditis symptom rate decreased by phage therapy, but not antibiotic group. The dosage of 0.1 mL per chicken in phage therapy dropped the rate from 53.1% to 16.3%; and the dosage of 0.3 mL per chicken, 43.3% to 21.8%; 0.5 mL per chicken, 44.8% to 11.7%. The antibiotic group was from 14.3% to 21.4%. The conclusion of this experiment was that the cocktail phage therapy could effectively control bacterial diseases in broiler feeding. However the dosage of phages was relevant in the therapy, and 0.1~0.3 mL per chicken was recommended, which equaled to 1~3×107 pfu/mL phage titer per chicken.
Defining the function of conserved hypothetical gene products encoded by an Stx phage
Hashim Felemban1,2, Alan J McCarthy1 and Heather E Allison1
1University of Liverpool, Institute of Integrative Biology, BioSciences Building, Crown Street, Liverpool, UK
2King Abdulaziz University, Faculty of Applied Medical Science, Jeddah, Saudi Arabia
Stx phages convert their bacterial hosts, providing them with shigatoxigenic potential. All E. coli carrying Stx phage are known as Shiga toxin-encoding Escherichia coli (STEC) have become a global challenge to food safety. Around 74% of the genes carried by the model Stx phage, ϕ24B (vb_EcoP-24B), are annotated as hypothetical, and we have been ascribing function to many of these genes. The expression of some hypothetical genes has been shown to be uncoupled from viral replication, but function to aid the lysogen in surviving environmental stresses such as antibiotic and acid exposures. Other genes, like gene 21, the target of this study, are expressed only during phage lytic replication. Gene 21 lies immediately upstream of the S, R, Rz and Rz1 genes. The gene product, P21, has been shown to possess an enzymatically active carbohydrate esterase domain (amino terminus), but its preferred biological substrate remains to be determined. We have created a series of isogenic mutants to establish whether P21 impacts upon phage release. New data show that P21 restricts the escape of 90% of progeny phages from their E. coli K12 hosts and limits the production of phage in an E. coli O157:H7 host, though release happens 10 min earlier. Work is currently ongoing to better understand how the action of P21 controls these phage release phenotypes. Peptidoglycan modification is used by Gram negative bacteria to increase their fitness and is a known phenomenon. Due to the genomic context and phenotype, our current hypothesis is that P21, a protein not encoded by lambda phage, alters the sensitivity of the lysogen’s peptidoglycan, altering the efficiency of the lytic burst. Why the phage would do this left to speculation, but could be due to the significant fitness advantage provided to the lysogen when the phage exists as a resident prophage.
Development of predictive phage therapy to treat multidrug resistant E. coli urinary tract infections in dogs and ultimately humans
Alison S Low, Nadejda Lupalova and David L Gally
Roslin Institute, Division of Infection & Immunity, University of Edinburgh, Easter Bush, EH25 9RG, UK
The rise of antibiotic resistant bacterial infections is a threat to the treatment of infectious diseases with companion animals acting as a potentially important reservoir in the transmission of resistant bacteria to and from humans. Our on-going research has been studying and sequencing multi-drug resistant (MDR) E. coli associated with urinary tract infections. Conventional treatments for these MDR infections are limited and we are developing the use of predictive phage therapy to efficiently select lytic phages that can be used for treatment. Although phage therapy has been used clinically in the former Soviet Union and Eastern Europe for decades, one limitation has been the time taken to find a suitable phage to treat a specific infection and this is especially true when applied to Gram-negative bacteria such as E. coli. We aim to develop predictive phage therapy based on collecting data from E. coli-phage interactions with high throughput methods. With this data, bioinformatics approaches and machine-learning, we will predict specific phage combinations based on the whole genome sequence of an isolate. Our initial proof of principal used E. coli O157:H7 as a test data set as whole genome sequences and phage sensitivity profiles (phage type) were available to analyse. With this data set we could predict phage sensitivities with test E. coli O157:H7 strains as well as identify specific genes present or absent when a strain was sensitive/resistant to a particular phage. We then tested the predictive capacity of the machine learning pipeline with sequenced canine MDR E. coli isolates using data collected by measuring interactions with 100 bacterial isolates and 2 phages using a 96-well plate reduction assay. The aim is to further test the predictive capacity of our system by collecting data from more phage-bacterial interactions and eventually the ambition is to produce bespoke phage cocktails that can be trialled to treat unresponsive UTIs.
Towards encapsulation of bacteriophages to improve effective delivery to the intracellular bacterial pathogen Rhodococcus equi
Anas Malas1, Alex Faulkner1, Martha Clokie1,. Natalie Garton1
Department of Infection, Immunity, and Inflammation, University of Leicester, University Road, Leicester, UK
Bacterial antibiotic resistance represents a significant challenge to global medicine. Bacteriophage (phage) therapy is receiving renewed attention as a promising alternative, or supplement to, conventional antibiotics and is being investigated for a number of important pathogens. However, bacteriophages do not penetrate eukaryotic cells well, limiting their use against intracellular bacterial pathogens. Liposome-encapsulation is a well-established method of improving the efficiency and targeting of drugs to sites of infection. Liposomes penetrate eukaryotic cells and can also be modified to tailor delivery to particular cell types. We aim to formulate and test liposome-encapsulated phage preparations for activity against Rhodococcus equi (Req) infections of macrophages. Req infection of foals has major economic impact on the equine breeding industry worldwide. Req inhabits soil and is transmitted in airborne particles. Req causes severe pneumonia in foals aged 1-6months which is often fatal if untreated. Antibiotic treatment is protracted and increasing incidence of resistant infections is being reported. An initial set of three Req phages (E3, V3 and V9) and 5 Req strains were assessed to establish a macrophage infection model and select an appropriate phage to take forward for liposome-encapsulation. The Req strains were screened for the virulence plasmid required for intracellular infection. Plasmid-containing Req 103S was then used to establish an infection model in murine J774.1 macrophages. Only phage V3 was found to produce a lytic infection Req 103S, however, infection occurred only at 30˚C and not 37˚C, which limits the potential of V3 in the treatment of infection in foals. No Req phages which produce lytic infection at 37˚C have been reported. Novel Req phages have been isolated from soil samples collected at studs with previous history of foal Req infection. These include phages which do produce lytic Req infection at 37˚C.
Isolation and characterization of new Campylobacter phages
Ibai Nafarrate and Amaia Lasagabaster
AZTI, Food Research Division, Parque Tecnológico de Bizkaia, Astondo bidea, Ed. 609, 48160 Derio, Bizkaia, Spain
Campylobacteriosis is the most commonly reported foodborne-illness in the European Union. Poultry is the natural reservoir of Campylobacter spp. and the consumption of chicken meat the most common route for human infection. The use of antimicrobial agents in food animal production has led to the spread of antibiotic resistant strains, an additional threat to public health. To reduce the burden of Campylobacter contamination within the farm-to-fork process, the use of Campylobacter specific bacteriophages has been suggested. In this study, twelve phages were isolated from pig feces and another one from chicken feces by enrichment in Bolton broth with a mix of 10 strains including C. jejuni, C. coli, C. lari and C. fetus. Regarding their host-range diversity, phages were classified into four lysis profiles. Phages of profile 1 exhibited the broadest host-range infecting more than the 80% of tested Campylobacter strains. Phages Cph100 and Cph047 lysed 9 from 10 strains, including 4 C. jejuni, 4 C. coli and 1 C. lari or 1 C. fetus, respectively, whereas Cph063 infected 8 from 10 strains, including 4 C. jejuni, 3 C. coli and 1 C. lari. Phages of profile 2 infected more than 70% of tested strains while those grouped in profiles 3 and 4 were more specific and only lysed 50 and 30% of tested strains, respectively. Phages of lysis profile 1, 2 and 3 presented a genome size of approximately 190 kb and could be, therefore, classified into group II or Cp220likeviruses. The phage of profile 4, instead, exhibited a genome size of 140 kb so it could be then categorized into the group III or Cp8unalikevirus.The broad host range of the 3 phages inducing lysis against more than 80% of tested Campylobacter isolates suggests their potential as biocontrol agents. However, further characterization is being carried out to determine their suitability for future development of new campylophage-based products.
Isolation of bacteriophages from environmental samples with lytic activity against Clostridium difficile
Supakan Panturat1, Sittinan Chanarat1, Surang Chankhamhaengdecha2
1Department of Biochemistry, Faculty of Science, Mahidol University, Thailand
2Department of Biology, Faculty of Science, Mahidol University, Thailand
Clostridium difficile is a significant cause of enteric diseases, including antibiotic-associated diarrhea and pseudomembranous colitis. Although antibiotics, metronidazole and vancomycin, are served as the primary treatment of C. difficile infection (CDI), the incidence and virulence of CDI still continuously increase due to the emergence of antibiotic resistant strains. The use of bacteriophages (phages) offers a possible alternative way for improving the infection treatment. In this study, we aimed at isolating phages with lytic activity against C. difficile from environments. Samples were collected from 29 different environmental sources in Thailand, including sewage water and soil contaminated with animal feces. After enrichment using 93 strains of C. difficile, we successfully isolated two C. difficile phages. One of them was purified and its stability at various temperatures and pH was investigated. Our results indicated the phage is most stable at pH 5-8 and 25°C. In addition, phage morphology was characterized under transmission electron microscope (TEM). The phages isolated in this study will help to develop a phage therapy for CDI in future.
Hydrogel as bacteriophage storage, assay and transportation matrix
Sheetal Patpatia1, Lauri Paasonen2, Mikael Skurnik1,3, Saija Kiljunen1
1Department of Bacteriology & Immunology, Res Programs Unit, Immunobiology, University of Helsinki, Helsinki, Finland
2UPM-Kymmene Corporation, Helsinki, Finland
3Division of Clinical Microbiology, Helsinki University Hospital, HUSLAB, Helsinki, Finland
Abstract excluded on presenter’s request
Characterization of fHyAci03, a novel lytic bacteriophage that infects clinical Acinetobacter strains
Elsi Pulkkinen2,1, Anu Wicklund2,1, Mikael Skurnik1,2, and Saija Kiljunen1
1Department of Bacteriology and Immunology, Medicum, Research Programs Unit, Immunobiology Research Program, University of Helsinki, Helsinki, Finland
2Division of Clinical Microbiology, HUSLAB, University of Helsinki and Helsinki University Hospital, Helsinki, Finland.
The genus Acinetobacter encompasses multiple nosocomial opportunistic pathogens. Among the genus, Acinetobacter baumannii, Acinetobacter pittii, and Acinetobacter nosocomialis are the most frequently isolated species in hospitals around the world. They have been implicated in a wide spectrum of infections, such as, bacteremia, secondary meningitis, ventilator-associated pneumonia, and urinary tract infection. Recently they have become a public health concern because of their growing trend for antibiotic resistance. We present here the isolation and characterization of Acinetobacter pittii phage vB_ApiM_fHyAci03 (fHyAci03), which is a new member of the T4-like virus family of Acinetobacter phages. It was isolated from municipal sewage sample collected in Finland. Phylogenetic tree for phage genome sequence revealed that fHyAci03 is closely related to Acinetobacter baumannii phage ZZ1 (HQ698922). The genome of phage fHyAci03 genome was found to be 165,975 bp in length and contain 255 genes, none of which encodes for toxins, virulence factors, antibiotic resistance, nor lysogeny. Host range experiments revealed that fHyAci03 could infect two pathogenic strains of A. nosocomialis, two pathogenic strains of A. baumannii, and six pathogenic strains of A. pittii. We conclude that fHyAci03 is a lytic phage that suits well to be used in phage therapy.
Isolation and characterization of new Flavobacterium columnare strains and its bacteriophages from fish farms
Anniina Runtuvuori1, Heidi Kunttu1, Gabriel MF Almeida1, Mathias Middelboe2, and Lotta-Riina Sundberg1
1Department of Biological and Environmental Science, University of Jyväskylä, Finland
2Department of Biology, Marine Biological Section, University of Copenhagen, Denmark
Fish are an important source of protein and aquaculture has been the fastest growing food production sector over the past 20 years. However, the intensification of fish production in farming systems expose fish to bacterial infections. Flavobacterium columnare is a Gram-negative bacterium that causes epidemical columnaris disease at fish farms during the warm water period. This pathogen causes significant financial losses in the industry if the disease is not treated with antibiotics as soon as the first symptoms appear. In order to develop a bacteriophage-based control of F. columnare, we isolated 117 F. columnare strains and 64 F. columnare phages from 10 different fish farms in Finland and Sweden. Genetic characterization of the bacterial isolates was made using 16S RFLP and ribosomal intergenic spacer analysis, and the virulence of 25 isolates was tested using a rainbow trout challenge model. All F. columnare isolates could be assigned into four previously identified genetic groups: 73 to group C, 22 to group E, 14 to group A and 8 to group G. Of these, bacterial isolates belonging to group C and E were the most virulent. Phage host range analysis against a collection of 229 bacterial isolates showed that the collection of phages together infected bacterial isolates from all the different genetic groups. Most phages infect bacteria in a genotype-specific manner, and only few isolates had wider host ranges. Phages infecting the most virulent bacterial groups were isolated. Together, the diversity, lytic capacity and combined host range of our phage collection demonstrates its potential to be used in phage therapy.
Bacteriophage efficacy against Staphylococcus aureus and Pseudomonas aeruginosa ex vivo isolates from diabetic foot ulcer (DFU) patients
Legesse Kifelew1, Morgyn Warner2, Sandra Morales3, Nicky Thomas6, David Gordon4, Robert Fitridge5, Peter Speck1
1College of Science and Engineering, Flinders University, South Australia
2Infectious Diseases Unit, The Queen Elizabeth Hospital, Woodville, South Australia
3AmpliPhi Biosciences Corporation, Brookvale, NSW, Australia
4College of Medicine and Public Health, Flinders University, South Australia
5Faculty of Health and Medical Sciences, University of Adelaide, South Australia
6School of Pharmacy and Health Science, University of South Australia
The global incidence of diabetes mellitus is rising, with ca. 5% of Australian adults (8% in the US) living with diabetes. A common complication of diabetes is foot ulcers, which become infected, often with S. aureus or P. aeruginosa. These bacterial species are frequently antibiotic resistant, so there may be failure to heal and resultant amputation of the foot. This procedure carries a 5-year mortality of >70%, worse than for many malignancies. Phages could provide a useful antimicrobial effect in this setting. We have access to phage cocktails AB-SA01, against S. aureus, and AB-PA01, against P. aeruginosa, both manufactured by AmpliPhi Biosciences. For phage cocktails to be useful in the DFU setting, minimum requirements to be met are: phages must be effective against their targets in biofilm form, and phages must be broadly effective against clinical isolates of their target species. To examine the potential efficacy of these phage products in the infected DFU setting, we obtained isolates of S. aureus (80 isolates) and P. aeruginosa (40 isolates) from infected DFUs in patients attending diabetic foot clinics in South Australia. The identity of bacterial isolates was confirmed by MALDI-TOF mass spectrometry. All isolates were tested for susceptibility to phage lysis by spotting serial dilutions of AB-SA01 or AB-PA01 onto bacterial lawns; 88% of S. aureus and 92% of P. aeruginosa isolates were lysed. Biofilms of target bacteria were grown on microtiter plates for 24hr, and phage cocktails then added. Biomass was measured at 24hr and 48hrs by crystal violet staining; 77% of S. aureus and 81% of P. aeruginosa isolates showed significant reduction in biomass when compared to untreated controls. This study indicates that phage cocktails AB-SA01 and AB-PA01 can potentially play a role in the treatment of diabetic foot ulcer infections.
Matching restriction modification systems to bacteriophages in Helicobacter pylori
Roxana Zamudio Zea, Liam A. Crawford, Andrew Millard, Sandra Beleza, Marco R. Oggioni
Department of Genetics and Genome Biology, University of Leicester, Leicester LE1 7RH, UK
The human pathogen Helicobacter pylori colonises approximately half of the global population and infection can lead to a range of gastric diseases. The temperate bacteriophages in H. pylori have been poorly characterised, most likely due to a very high number and strain-to-strain variability of restriction modification (RM) systems, which can be easily more than 20 in any strain. This work aims to study the prevalence of bacteriophages and RM systems in over 460 strains of H. pylori isolated from 184 gastric samples from asymptomatic subjects. H. pylori prophages were identified using the PHASTER tool. The gold standard RM systems were downloaded from Rebase and the RM genes were identified in the 460 genomes using Blast+. The analysis for phage genomes showed 57 intact bacteriophages, ranging from 12 – 30 Kb in length, in 25 subjects (14%). Approximately half of the bacteriophages were shown to have integrated in the Lipid A biosynthesis gene lpxD. Using 107 RM system genes, we built a presence/absence matrix of the genes in all our H. pylori genomes, which was ordered according to a maximum likelihood gene presence/absence tree generated by FastTree. This clustering revealed the presence of multiple pairs of strains of compatible RM systems, one of which carrying an intact phage (potential phage donor) and one an intact target site (potential phage recipient), five of which with a perfect match of RM systems. The availability of this panel of donor and recipient strain pairs is an ideal starting point to study the molecular biology of bacteriophages in H. pylori.
