Ana Rodríguez González
Ana Rodríguez González has a Bachelor’s degree in Biological Sciences (specialisation: Microbiology, Biochemistry and Genetics) from the Universidad de Oviedo (1979) and a PhD from the same university (1984), having completed her doctoral thesis in the field of microbiology. She subsequently realised a postdoctoral fellowship (1985-1987) in the Genetics and Microbiology Department of the AFRC Institute of Food Research (Reading Laboratory, Reading, UK). In 1988, she joined the staff of CSIC’s Instituto de Productos Lácteos de Asturias (IPLA-CSIC) in Villaviciosa, where she was Deputy Director from 1995 to 2009.
Pilar García Suárez
Pilar García Suárez has a Bachelor’s degree in Biology from the Universidad de Oviedo (1988) and a PhD from the same university (1993), having completed her doctoral thesis in the field of microbiology. In 2005, she joined IPLA-CSIC’s staff on a Ramón y Cajal contract. She obtained the position of CSIC Research Scientist in 2009, and she is currently Director of the Biotechnology and Dairy Product Technology Department.
|Ana Rodríguez González||PRO (Public Research Organisation) Senior Research Scientist|
|Pilar García Suárez||PRO Research Scientist|
Principal Investigator on Projects and Director of the Biotechnology and Dairy Product Technology Department
|Beatriz Martínez Fernández||PRO Senior Research Scientist|
Principal Investigator on Projects
|Lucía Fernández Llamas||Postdoctoral Researcher|
Grants for Research Groups in the Principado de Asturias Project (2018-2020). Ref. IDI/2018/000119
|Diana Gutiérrez Fernández||Postdoctoral Researcher|
EU ERA-NET Projects – CSIC Intramural Project Ref. 201670E040
|Susana Escobedo Martín||Postdoctoral Researcher|
|Rosana Calvo Méndez||Technical and Professional Officer|
|Eva González Menéndez|
|Silvia González Menéndez|
|María Jesús López González|
The DairySafe (Lactic Ferments and Biopreservation) research group has been closely connected to IPLA-CSIC since the group was created (1990), Dr Ana Rodríguez González having been its coordinator since then.
Biopreservation has been the group’s main line of research over the past 20 years, with a particular focus on developing food preservation strategies based on natural antimicrobials that improve food safety. The antimicrobials studied include bacteriocins produced by lactic acid bacteria, and bacteriophages that infect food pathogens, as well as phage-synthesised lytic proteins . The most significant objectives of this work include exploring the mode of action of these antimicrobials, the consequences of their use, and the possible resistance mechanisms of the target bacteria.
Over this period, we have isolated and characterised bacteriocin-producing lactic bacteria (nisin Z, lacticin 481, coagulin A and lactococcin 972). Producer strains of nisin Z act effectively as protective cultures in cheese production, while lactococcin 972 inhibits cell wall synthesis in lactococci through interaction with lipid II (an intermediate in peptidoglycan synthesis) and generates the two-component CesSR system which controls the cell wall’s immediate response to stress in Lactococcus lactis. Lactococcin 972 is being used in the adaptive evolution of lactococci to obtain more robust strains with improved technological aptitudes that are useful for the food industry. Coagulin A has a high level of activity against Listeria monocytogenes.
Bacteriophages, a natural enemy of bacteria, are a promising alternative to the use of chemical compounds such as biopreservatives and disinfectants in the food industry. The specificity of bacteriophages, which are active against one bacterial species and even against certain strains only, makes them completely harmless to humans, animals and the environment. In light of these insights, our group has been pioneering: the use of phage mixtures against Staphylococcus aureus in milk, curd and cheese; the combined use of phages and bacteriocins in milk to inhibit S. Aureus, and detecting the activity of recombinant phage endolysin in milk. It is worth noting that S. aureus is responsible for a wide range of infections in humans and animals, including toxic food infections caused by the consumption of food contaminated by enterotoxins. Phages and phage-synthesised lytic proteins are effective against S. aureus strains of human and animal origin – even those that are resistant to antibiotics – as well as against Staphylococcus epidermidis. They are equally effective whether the strains are biofilm producers or not. In fact, our group has characterised several phage-derived proteins that have lytic activity against biofilm producers, as well as a protein with polysaccharide depolymerase activity that breaks down the extracellular matrix of biofilms. The group has also characterised the phenotypic and transcriptional responses of biofilms exposed to low doses of bacteriophages and lytic proteins, in order to optimise their doses for different applications.
Our results clearly demonstrate the great potential of bacteriophages and their lytic proteins as alternatives to current antimicrobials and disinfectants for tackling S. aureus and S. Epidermidis infections and contamination, with applications in clinical and veterinary settings, as well as the food industry.
We have recently validated an impedance-based method for monitoring the ability of various pathogenic bacteria to form biofilm (Real Time Cell Analyser, RCTA). The method is quick, reliable and easy to use for several purposes, including analysing biofilm-producing strains, and searching for the antimicrobial compounds which inhibit the formation of this type of structure or which eliminate those that have already been formed.
Current and Future Priorities
Taking the knowledge acquired by the DairySafe group on new antimicrobials as a starting point, our main objective is to use these antimicrobials to eliminate pathogens that are of concern to hospitals. In this context, and due to the urgent need for alternatives to antibiotics, we intend to continue expanding our knowledge of the mechanisms that promote the action of new antimicrobials against antibiotic-resistant bacteria, and also against biofilm producers, given that these structures’ powerful protective effect makes them more difficult to eliminate. Our aim is to develop effective antimicrobials against this type of bacteria, as well as detection systems that enable early diagnosis of infections.
The group has many years of experience in developing food preservation strategies based on natural antimicrobials (biopreservation) that improve food health and hygiene standards, thereby reducing the frequency of foodborne outbreaks. Of these antimicrobials, we have paid particular attention to the bacteriocins produced by lactic acid bacteria (LAB), the bacteriophages that infect food pathogens, and phage-synthesised lytic proteins (endolysins).
Our work on bacteriocins has included designing protective cultures made up of producer strains, which have been successfully used in the production of dairy products.
Natural Antimicrobials: Genetic Regulation and Function
Understanding the mode of action of antimicrobial substances enables us to learn about the molecular mechanisms by which bacteria become resistant to them. Using the Lcn972 bacteriocin as a model, the group has studied both the mode of action and gene expression. This bacteriocin inhibits cell division: a completely different mode of action to the other bacteriocins produced by LAB. Lcn972 is the first bacteriocin described that interacts with Lipid II, without having been modified after transduction. It also generates the two-component CesSR system, which controls the cell wall’s immediate response to stress in Lactococcus lactis. Lnc972 is being used to study the adaptive evolution of L. lactis under conditions of cell-wall stress. This food-grade strategy is focused on obtaining more robust strains with improved technological aptitudes that are useful for the food industry, as well as strains that can be used as cell factories in the production of enzymes with clinical and food applications.
Phage Therapy and Biocontrol
Bacteriophages and their endolysins are a promising alternative to the use of biocides, including preservatives and disinfectants (food industry) and antibiotics (health sector).
Bacteriophages specifically inhibit bacteria, making them completely harmless to other living beings and the environment. Our group has therefore been pioneering the use of phages and endolysins against Staphylococcus aureus, with the aim of tackling toxic food infections and health-related infections caused by this bacterium. Our phages and some phage-synthesised proteins (endolysins and virion-associated peptidoglycan hydrolases) are also effective against staphylococcus strains of human and animal origin, including Staphylococcus epidermidis and even antibiotic-resistant strains, whether they are biofilm producers or not.
The results obtained by our group clearly demonstrate the great potential of phage-based products for future applications in the clinical, veterinary and food sectors.
- Gutiérrez, D., Adriaenssens, E.M., Martínez, B., Rodríguez, A., Lavigne, R., Kropinski, A.M., García, P. 2014. Three proposed new bacteriophage genera of staphylococcal phages: “3alikevirus”, “77likevirus” and “Phietalikevirus”. Archives of Virology 159: 389-398 (doi 10.1007/s00705-013-1833-1). (A) (Q3). IF: 2.390
- Campelo, A.B., Roces, C., Mohedano, M.L., López, P., Rodríguez, A. and Martínez, B 2014. A bacteriocin gene cluster able to enhance plasmid maintenance in Lactococcus lactis. Microbial Cell Factories 13: 77. (Q1). IF: 4.221
- Diana Gutiérrez, Patricia Ruas-Madiedo, Beatriz Martínez, Ana Rodríguez and Pilar García. (2014). Effective removal of staphylococcal biofilms by the endolysin LysH5. PLoS ONE 9 (9):e107307 (Q1). IF: 3.234
- Rehaiem, A., Ben Belgacem, Z., Edalatian M.R., Martínez, B., Rodríguez, A., Manai, M. and Guerra, N.P. (2014). Assessment of potential probiotic properties and multiple bacteriocin encoding-genes of the technological performing strain Enterococcus faecium Food Control (doi:10.1016/j.foodcont.2013.09.044) (Q1) IF: 2.806
- Rodríguez-Rubio, L., García, P., Rodríguez, A., Billington, C., Hudson, J.A. and Martínez, B. (2015). Listeriaphages and coagulin C23 act synergistically to kill Listeria monocytogenes in milk under refrigeration conditions. International Journal of Food Microbiology 205: 68-72. doi: 10.1016/j.ijfoodmicro.2015.04.007 (Q1) IF: 3.445
- Milioni, B. Martínez, S. Degl’Innocenti, B. Turchi, F. Fratini, D. Cerri, R. Fischetti. (2015). A novel bacteriocin produced by Lactobacillus plantarum LpU4 as a valuable candidate for biopreservation in artisanal raw milk cheese. Dairy Science and Technology 95: 479–494 (Q3) IF: 1.435
- Diana Gutiérrez, DieterVandenheuvel, Beatriz Martínez, Ana Rodríguez, Rob Lavigne and Pilar García. Two phages, phiIPLA-RODI and phiIPLA-C1C, lyse mono- and dual-staphylococcal biofilms. Applied and Environmental Microbiology 81: 3336-3348. doi:10.1128/AEM.03560-14. (Q1) IF: 3.823
- Diana Gutiérrez, Yves Briers, Lorena Rodríguez-Rubio, Beatriz Martínez, Ana Rodríguez, Rob Lavigne and Pilar García. (2015). Role of the pre-neck appendage protein (Dpo7) from phage vB_SepiS-phiIPLA7 as an anti-biofilm agent in staphylococcal species. Frontiers in Microbiology 6: 1315 (doi: 10.3389/fmicb.2015.01315) (Q1) IF: 4.165
- Roces, C., Campelo, A.B., Escobedo, S., Wegmann, U., García, P., Rodríguez, A. and Martínez, B. (2016). Reduced binding of the endolysin LysTP712 to Lactococcus lactis ΔftsH contributes to phage resistance. Frontiers in Microbiology 7: 138 (doi: 10.3389/fmicb.2016.00138) (Q1) IF: 4.076
- Lorena Rodríguez-Rubio, Diana Gutiérrez, David M. Donovan, Beatriz Martínez, Ana Rodríguez and Pilar García. (2016). Phage lytic proteins: biotechnological applications beyond clinical antimicrobials. Critical Reviews in Biotechnology 36 (3): 542-552. (doi:10.3109/07388551.2014.993587). (Q1) IF: 6.542
- Gutiérrez, D., Rodríguez-Rubio, L., García, P., Billington, C., Premarante, A., Rodríguez, A. and Martínez, B. (2016). Phage sensitivity and prophage carriage in Staphylococcus aureus isolated from foods in Spain and New Zealand. International Journal of Food Microbiology 230: 16-20. (Q1) IF: 3.339
- Gutiérrez D, Rodríguez-Rubio L, Martínez B, Rodríguez A and García P (2016) Bacteriophages as Weapons against Bacterial Biofilms in the Food Industry. Frontiers in Microbiology. 7: 825. (doi: 10.3389/fmicb.2016.00825) (Q1) IF: 4.076
- Lorena Rodríguez-Rubio, Wai-Ling Chang, Diana Gutiérrez, Rob Lavigne, Beatriz Martínez, Ana Rodríguez, Sander K. Govers Abram Aertsen, Christine Hirl, Manfred Biebl, Yves Briers, Pilar García. (2016). ‘Artilysation’ of endolysinλSa2lys strongly improves its enzymatic and antibacterial activity against streptococci. Scientific Reports 6: 35382. doi: 10.1038/srep35382. (Q1) IF: 4.259
- Diana Gutiérrez, Claudio Hidalgo-Cantabrana, Ana Rodríguez, Pilar García, Patricia Ruas-Madiedo (2016). Monitoring in Real Time the Formation and Removal of Biofilms from Clinical Related Pathogens Using an Impedance-Based Technology. PLoS ONE 11 (8): e0163966 doi:10.1371/journal.pone.0163966. (Q1) IF: 2.806
- Diana Gutiérrez, Lorena Rodríguez-Rubio, Lucía Fernández, Beatriz Martínez, Ana Rodríguez, Pilar García. (2017). Applicability of commercial phage-based products against Listeria monocytogenes for improvement of food safety in Spanish dry-cured ham and food contact surfaces. Food Control 73: 1474-1482. http:// dx.doi.org /10.1016/j.foodcont.2016.11.007. (Q1) IF: 3.496
- Silvia González, Lucía Fernández, Ana Belén Campelo, Diana Gutiérrez, Beatriz Martínez, Ana Rodríguez, Pilar García. (2017). The behavior of Staphylococcus aureus dual-species biofilms treated with bacteriophage phiIPLA-RODI depends on the accompanying microorganism. Applied and Environmental Microbiology 83(3): e02821. (Q1) IF: 3.807
- Abedon, S.; García, P.; Mullany, P. and Aminov, R. (2017). Editorial: Phage therapy: past, present and future. Frontiers in Microbiology. 8: 981. doi: 10.3389/fmicb.2017.00981 (Q2) IF: 4.076
- Lucía Fernández, Silvia González, Ana Belén Campelo, Beatriz Martínez, Ana Rodríguez and Pilar García. (2017). Low-level predation by lytic phage phiIPLA-RODI promotes biofilm formation and triggers the stringent response in Staphylococcus aureus. Scientific Reports 7, 40965; doi: 10.1038/srep40965 (Q1) IF: 4.259
- Gutiérrez D.; Fernández L.; Martínez B.; Ruas-Madiedo P., García P.; Rodríguez A. (2017). Real-Time Assessment of Staphylococcus aureus Biofilm Disruption by Phage-derived Proteins. Frontiers in Microbiology 8: 1632 (Q2) IF: 4.076
- Fernández, L.; González, S; Campelo, A.B.; Martínez, B.; Rodríguez, A. and García, P. (2017). Downregulation of autolysin-encoding genes by phage-derived lytic proteins inhibits biofilm formation in Staphylococcus aureus. Antimicrobials Agents and Chemotherapy 61 (5): e02724-16. https://doi.org/10.1128/AAC.02724-16 (Q1) IF: 4.302
|Principal Investigator||Title||Funding Body||Reference No||Duration||Researchers|
|Beatriz Martínez||Structure and function of bacteriocin Lcn972: determination of binding to Lipid II domain and target cells||MCINN||BIO2010-17414||2011 – 2013||Ana Rodríguez|
|Beatriz Martínez||Increasing dairy productivity and food safety using biocontrol||MINECO: joint action with New Zealand||PRI-AIBNZ-2011-1043||2011 – 2013||Ana Rodríguez|
|Juan C. Alonso||FAGOMA (Spanish Network of Bacteriophages and Transducing Elements)||MICINN||BFU2010-10469-E||2011 – 2012||Pilar GarcíaBeatriz Martínez|
|Jan Wedekind (IRIS Spain)||Smartmilk: a novel system for the treatment of milk, based on the combination of ultrasounds and pulse electric field technologies||EU 7th Framework Programme (SME Project)||FP7-261591||Oct 2010 – Mar 2013||Ana Rodríguez|
|Fernando Vicente (SERIDA: Regional Agri-Food Research and Development Service of Asturias)||’Small-scale’ milk production as a tool for boosting economic development in the Central Mexican Plateau||Cooperation Project: AECID (Spanish Agency for International Development Cooperation) – DGPOLDE (General Office for Planning and Evaluation of Development Policies)||AECID11-CAP2-1526||2011 – 2013||Ana Rodríguez|
|Pilar García||Control of mixed biofilms of Staphylococcus aureus in the food industry by using bacteriophages and phage-derived proteins||MICINN||AGL2012-40194-C02-01||2013 – 2015||Ana Rodríguez|
|Beatriz Martínez||New biopreservatives for the food industry||Scientific partnership project for developing i-COOP cooperation (Spain-Mexico)||COOPA200015||2014 – 2015||Ana Rodríguez|
|Beatriz Martínez||Improvement of the biotechnological yield of Lactococcus lactis by modulating the cellular wall||MINECO||BIO2013-46266-R||2014 – 2017||Ana Rodríguez|
|Ana Rodríguez||Research teams funding: DairySafe Team||Science, Technology and Innovation Programme 2013-2017, Principado de Asturias, Spain||GRUPIN14-139||2014 – 2017||Beatriz MartínezPilar García|
Ana B. Campelo
Juan E. Suárez
|Pilar García||FAGOMA II (Spanish Network of Bacteriophages and Transducing Elements II)||MINECO||BIO2015-70394-REDT||2015 – 2018||Ana RodríguezBeatriz Martínez|
|Ana Rodríguez||Bacteriophage lysins as alternatives to antimicrobial treatment||EU ERA-NET||BLAAT ID: 67||2016 – 2018||Pilar GarcíaBeatriz Martínez|
|Pilar García||Bacteriophages and phage proteins: a step towards their application in the food industry||Ministerio de Economía, Industria y Competitividad||AGL2015-65673-R||2016 – 2018||Ana Rodríguez|
|Ana Rodríguez||Phage endolysins as biopreservatives and/or disinfectants for the food industry as an alternative to antibiotic treatment||Intramural CSIC Project||201670e040||April 2016 – March 2019||Diana Gutiérrez|
|Ana Rodríguez||Study of the bacterial response of biofilms treated with bacteriophages||Intramural CSIC Project||201770e016||January 2017 – December 2018||Lucía Fernández|
|Beatriz Martínez||Functional analysis of the cell envelope stress response in Lactococcus lactis and its consequences for bacteriophage infection||Ministerio de Economía, Industria y Competitividad||BIO2017-88147-R||2018 – 2020||Ana Rodríguez|
|Ana Rodríguez||Research teams funding: DairySafe Team (Ref. ) (): PI:. Researchers:||Science, Technology and Innovation Programme 2018-2020, Principado de Asturias, Spain||IDI/2018/000119||2018-2020||Beatriz Martínez FernándezPilar García Suárez|
Lucía Fernández Llamas
Diana Gutiérrez Fernández
Lucía Escobedo Martín.
|Supervisors||Title||PhD Student||Organisation||Date of Viva||Classification|
|Beatriz Martínez FernándezAna Rodríguez González||Molecular response mechanisms to stress applied to the cell wall in Lactococcus lactis||Clara Roces Rodríguez||Faculty of Biology, Universidad de Oviedo.||08/02/2013||Cum LaudeEuropean Honours.|
|Pilar García SuárezAna Rodríguez González||The potential of phages and phage proteins for eliminating biofilms formed by Staphylococci||Diana Gutiérrez Fernández||Faculty of Biology, Universidad de Oviedo||8/5/2015||DistinctionInternational Honours|
|Patricia G.Mendoza GarcíaMario Ramírez Lepe|
Beatriz Martínez Fernández
|Isolation and characterisation of bacteriocin-producing lactic acid bacteria in regional artisan cheeses||Silvia Portilla Vázquez||Instituto Tecnológico de Veracruz (Veracruz Technological Institute), Mexico||15/06/2015||Pass (unanimous)|
- Enhanced staphylolytic activity of the Staphylococcus aureus bacteriophage vB_SauS-phiIPLA88 HydH5 virion associated peptidoglycan hydrolase: fusions, deletions and synergy with LysH5
Inventors: David M. Donovan, Lorena Rodríguez-Rubio, Beatriz Martínez, Ana Rodríguez, Pilar García.
Application No: US13298966
Patent Assignment: 501810442 (6 February 2012)
Priority Country: US
Owner: CSIC – USDA (United States Department of Agriculture)
Priority Date: 17/11/2011
US Patent No 8,986,695, issued on 24/03/2015
- Chimeric protein with high antimicrobial activity
Inventors: Diana Gutiérrez, Beatriz Martínez, Pilar García, Ana Rodríguez, Lorena Rodríguez-Rubio, Rob Lavigne
Application No: PCT/EP2018/082715 (27 November 2018)
Reference Number: EP1641.1323