Projects / Programmes source: ARRS

Development of bacteriophage characterization platform to support their use as an antibacterial agents

Research activity

Code Science Field Subfield
4.06.04  Biotechnical sciences  Biotechnology  Microbe biotechnology 

Code Science Field
T490  Technological sciences  Biotechnology 

Code Science Field
2.08  Engineering and Technology  Environmental biotechnology  
bacteriophages, bacteriophage therapy, Campylobacter, cell culture model, immune response, Campylobacter
Evaluation (rules)
source: COBISS
Researchers (17)
no. Code Name and surname Research area Role Period No. of publications
1.  12667  PhD Miloš Barut  Chemistry  Researcher  2013  206 
2.  29430  PhD Ivanka Cizelj  Biotechnology  Researcher  2014 - 2016  51 
3.  28558  PhD Jernej Gašperšič  Biochemistry and molecular biology  Researcher  2016  62 
4.  33406  PhD Nikolaja Janež  Biochemistry and molecular biology  Researcher  2013 - 2016  66 
5.  29864  Marko Jarc  Biotechnology  Researcher  2014 - 2015  22 
6.  22491  PhD Anja Klančnik  Animal production  Researcher  2013 - 2016  315 
7.  35145  Maja Leskovec  Biotechnology  Researcher  2016 
8.  05008  PhD Mojca Narat  Biotechnology  Researcher  2013 - 2016  675 
9.  24298  PhD Irena Oven  Animal production  Researcher  2013 - 2016  81 
10.  16327  PhD Matjaž Peterka  Biotechnology  Principal Researcher  2013 - 2016  215 
11.  12728  PhD Aleš Podgornik  Chemical engineering  Researcher  2013 - 2016  680 
12.  04001  PhD Peter Raspor  Microbiology and immunology  Researcher  2013  1,877 
13.  07030  PhD Sonja Smole - Možina  Animal production  Researcher  2013 - 2016  1,058 
14.  27846  PhD Lidija Urbas  Pharmacy  Researcher  2013 - 2015  49 
15.  36980  Urška Vidic  Biotechnology  Researcher  2016  28 
16.  36120  Eva Zaletel  Biotechnology  Technician  2014 - 2015  16 
17.  03683  PhD Mirjan Žorž  Chemistry  Researcher  2014 - 2015  128 
Organisations (3)
no. Code Research organisation City Registration number No. of publications
1.  0481  University of Ljubljana, Biotechnical Faculty  Ljubljana  1626914  64,462 
2.  1655  Sartorius BIA Separations L.t.d., Separations technology company  Ajdovščina  1319612  698 
3.  3030  Center of Excellence for Biosensors, Instrumentation and process Control  Ajdovščina  3660460  899 
Excessive use of antibiotics in the past decades increased incidence of antibiotic resistant pathogenic bacteria to the level that it become a significant problem. Only in Europe at least 25,000 patients die per year because their bacterial infections are not treatable with available antibiotics. New antibiotics and novel, sustainable antimicrobial strategies are needed. Bacteriophage therapy is one of the most verified approaches and has been applied for decades as a means of treating bacterial infections in Russia, Poland, and Georgia. Due to increase of antibiotic resistant interest spread to Western world and an increasing number of publications, as well as the emergence of bacteriophage companies indicates an increasing appetite among the scientific community and industry for developing bacteriophage therapy as part of antibacterial mainstream in medicine and food production. Very few human clinical trials have been conducted in either EMEA or FDA jurisdictions. These trials, alongside earlier, less well reported studies, indicate that phage therapy shows considerable promise for treating infectious bacterial diseases and as an alternative antibacterial agent in food production and industry. Although, knowledge of the biology, genetics and bactericidal efficacy of bacteriophages in vitro are impressive, less is known about their behavior in complex and dynamic interactions with bacterial host cell. Fully controlled experiments on molecular level as well as clinical trials need to be performed in order to develop robust, reliable data to answer the key efficacy and safety questions of bacteriophage therapy. Laboratory for bio-analytics of CO BIK is focused on bacteriophage based antimicrobials to control contamination in food chain and to treat some bacterial infections in humans. One of our goals is to prepare bacteriophage product against Campylobacter in different formulations applicable to animals, food and humans. Bacteria of the genus Campylobacter are most frequent agents responsible for human enteric diseases in Europe and developed world in general. While efficacy of bacteriophages against Campylobacter was already confirmed on animals, no human trial was performed until today. We presume that bacteriophages are effective against Campylobacter also in humans and such therapy can shorten the disease and the time bacteria are excreted by the infected person. As a first step towards clinical trials we will develop human cell culture based model to demonstrate bacteriophage efficacy against Campylobacter. Such model will enable us to monitor bacteriophage efficacy and immune response of human cells against bacteriophage and to study potential anti-inflammatory effect of bacteriophages. We will also develop methods for rapid identification of Campylobacter structures involved in the bacteria-phage interactions. Using these methods we will be able to characterize interaction and to monitor changes on these structures on bacteriophage resistant bacteria isolated from cell culture model. To perform these studies pure bacteriophage formulation is required. Therefore, we will develop purification methods to prepare GMP-like bacteriophage formulation applicable also on large scale production level. The main goal of the project is therefore biological characterization platform for Campylobacter bacteriophages to support their use as antibacterial agents. Developed platform will be applicable to other bacteria-phage systems. In the frame of the project we will: · Develop in-vitro human cell culture based model to determine efficacy of bacteriophages on adhered Campylobacter bacteria and influence on human cell line immune response · Develop a technology for rapid identification of receptor structures and monitoring of changes on bacterial hosts during bacteriophage therapy · Develop a chromatographic method for purification of bacteriophages to prepare GMP-like phage formulation
Significance for science
The increase in resistance to antibiotics almost all important pathogenic bacteria has moved the focus of research in the search for new antibiotics and antibacterial strategies. Bacteriophage therapy is one of the alternative approaches which are recognized and accepted in some parts of the world. In Europe and the USA the use of bacteriophages as antibacterial agents is known and used partially, but not fully accepted. During last years several clinical trials were completed, and some are ongoing. At the same time number of scientific publications that describe the use of bacteriophages as antibacterial agents is increasing. Knowledge of biology, genetics and antibacterial efficacy of bacteriophages is considerable but we know much less about their behavior in complex and dynamic society with bacterial hosts. To obtain the relevant data that would confirm the safety and efficacy of bacteriophages studies on the molecular level, as well as clinical studies conducted according to established standards are necessary. Within the project, we have developed a model that allows evaluating the efficacy and safety of bacteriophage. We have obtained data on the efficacy of isolated bacteriophages against Campylobacter. At the same time, we also gained insight on the impact of bacteriophages on human cells. This important safety aspect of the use of bacteriophages against Campylobacter has not yet been examined. Model allows, unlike an animal model that is in the world already known and established, the isolation of the target bacteria and bacteriophages and study of interactions between them. We have identified receptors at the molecular level and we monitored changes in these receptors during bacteriophage therapy. Data indicate the similarity of mechanisms of resistance in the system described above and an animal model. The methodology is transferable to other phage-bacterial systems and can become part of the platform for the characterization of bacteriophages, which will speed up the transfer of the bacteriophage products from preclinical to clinical studies.
Significance for the country
The resistance of bacteria to antibiotics is a global problem and the development of new antibiotics and their alternatives is one of the development priorities, which is also important for Slovenia. Results of the project put Slovenia strongly on the global map of the fight against bacteria resistant to antibiotics. Implementation of the project has strengthen cooperation with research groups from abroad, and established new connections and thereby increase the potential of research group on international calls for grants. Project results will enable the continuation of work towards a more rapid and efficient discovery of bacteriophages and bacteriophage enzymes. The project has encouraged bachelor, masters and research works and thus contributed significantly to the education of young researchers and expand awareness of the need for new antibacterial agents between the professional and the public. Results of the project have encouraged cooperation between research group with a company that is developing a new product, expanding set of products, and therefore plans new jobs with higher added value. A partner company expanded service at the global market, and preserve and enable jobs with high added value.
Most important scientific results Annual report 2013, 2014, 2015, final report
Most important socioeconomically and culturally relevant results Annual report 2013, 2014, 2015, final report
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