Research Divisions

Lactic Ferments and Biopreservation

Group LeadersLocation and ContactMembersHistory of the GroupLines of ResearchNotable PublicationsProjectsPhD ThesesPatents and TrademarksNews
Dr Ana Rodríguez

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.

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The DairySafe Group was established in 1990 under her supervision, its first line of research being the development of starter cultures for autochthonous cheeses, to which research on biopreservation was added, centred on developing food preservation strategies based on natural antimicrobials (bacteriocins produced by lactic acid bacteria). Dr Pilar García Suárez’s incorporation into the group in 2005 enabled a line of research to be initiated on biopreservation using bacteriophages and phage-synthesised lytic proteins.

She has been principal investigator on 13 projects under the Spanish National RDI Plan and the Science, Technology and Innovation Plan for the Principado de Asturias. Additionally, she has acted as IPLA-CSIC’s lead researcher on two joint research projects as part of the Ibero-America Cooperation Programme, one SME project in the EU’s 7th Framework Programme, and one ERA-Net ANIWHA project (EU). She has also participated as a researcher in 10 of the research group’s projects. She has supervised and/or co-supervised 10 PhD theses, and her research work has resulted in 114 SCI-listed articles, 20 chapters in national and international books and 25 non-SCI-listed and science communication articles.

Contact: anarguez@ipla.csic.es

 

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.

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Her research focuses on studying bacteriophages as tools for developing new antimicrobial agents that may be used for food safety. She has participated in 28 regional, national and international projects, acting as principal investigator in nine of them, and has led on four research contracts with international companies. She has supervised seven PhD theses and her research results have been published in over 100 scientific articles and book chapters.

Contact: pgarcia@ipla.cisc.es

Dr Pilar García

Location

Instituto de Productos Lácteos de Asturias (IPLA-CSIC)

Paseo río Linares s/n.

33300. Villaviciosa Asturias, Spain.

Contact

Ana Rodríguez González

+34 985893246

anarguez@ipla.csic.es

Pilar García Suárez

+34 985893420

pgarcia@ipla.csic.es

Beatriz Martínez Fernández

+34 985892233

bmf1@ipla.csic.es

Permanent Members

Ana Rodríguez GonzálezPRO (Public Research Organisation) Senior Research Scientist
Pilar García SuárezPRO Research Scientist
Principal Investigator on Projects and Director of the Biotechnology and Dairy Product Technology Department
Beatriz Martínez FernándezPRO Senior Research Scientist
Principal Investigator on Projects
Fixed-Term Staff
Lucía Fernández LlamasPostdoctoral 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 MartínPostdoctoral Researcher
BIO2017-88147-R Project
Rosana Calvo Méndez Technical and Professional Officer
AGL2015-65673-R Project
PhD Students
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.

Food Safety

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 InvestigatorTitleFunding BodyReference NoDurationResearchers
Beatriz MartínezStructure and function of bacteriocin Lcn972: determination of binding to Lipid II domain and target cellsMCINNBIO2010-174142011 – 2013Ana Rodríguez
Beatriz MartínezIncreasing dairy productivity and food safety using biocontrolMINECO: joint action with New ZealandPRI-AIBNZ-2011-10432011 – 2013Ana Rodríguez
Juan C. AlonsoFAGOMA (Spanish Network of Bacteriophages and Transducing Elements)MICINNBFU2010-10469-E2011 – 2012Pilar GarcíaBeatriz Martínez

Ana Rodríguez

Jan Wedekind (IRIS Spain)Smartmilk: a novel system for the treatment of milk, based on the combination of ultrasounds and pulse electric field technologiesEU 7th Framework Programme (SME Project)FP7-261591Oct 2010 – Mar 2013Ana 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 PlateauCooperation Project: AECID (Spanish Agency for International Development Cooperation) – DGPOLDE (General Office for Planning and Evaluation of Development Policies)AECID11-CAP2-15262011 – 2013Ana Rodríguez
Pilar GarcíaControl of mixed biofilms of Staphylococcus aureus in the food industry by using bacteriophages and phage-derived proteinsMICINNAGL2012-40194-C02-012013 – 2015Ana Rodríguez
Beatriz MartínezNew biopreservatives for the food industryScientific partnership project for developing i-COOP cooperation (Spain-Mexico)COOPA2000152014 – 2015Ana Rodríguez
Beatriz MartínezImprovement of the biotechnological yield of Lactococcus lactis by modulating the cellular wallMINECOBIO2013-46266-R2014 – 2017Ana Rodríguez
Ana RodríguezResearch teams funding: DairySafe TeamScience, Technology and Innovation Programme 2013-2017, Principado de Asturias, SpainGRUPIN14-1392014 – 2017Beatriz MartínezPilar García

Ana B. Campelo

Juan E. Suárez

Lucía Fernández

Diana Gutiérrez

Pilar GarcíaFAGOMA II (Spanish Network of Bacteriophages and Transducing Elements II)MINECOBIO2015-70394-REDT2015 – 2018Ana RodríguezBeatriz Martínez
Ana RodríguezBacteriophage lysins as alternatives to antimicrobial treatmentEU ERA-NETBLAAT ID: 672016 – 2018Pilar GarcíaBeatriz Martínez

Diana Gutiérrez

Pilar GarcíaBacteriophages and phage proteins: a step towards their application in the food industryMinisterio de Economía, Industria y CompetitividadAGL2015-65673-R2016 – 2018Ana Rodríguez
Ana RodríguezPhage endolysins as biopreservatives and/or disinfectants for the food industry as an alternative to antibiotic treatmentIntramural CSIC Project201670e040April 2016 – March 2019Diana Gutiérrez
Ana RodríguezStudy of the bacterial response of biofilms treated with bacteriophagesIntramural CSIC Project201770e016January 2017 – December 2018Lucía Fernández
Beatriz MartínezFunctional analysis of the cell envelope stress response in Lactococcus lactis and its consequences for bacteriophage infectionMinisterio de Economía, Industria y CompetitividadBIO2017-88147-R2018 – 2020Ana Rodríguez
Ana RodríguezResearch teams funding: DairySafe Team (Ref. ) (): PI:. Researchers:Science, Technology and Innovation Programme 2018-2020, Principado de Asturias, SpainIDI/2018/0001192018-2020Beatriz Martínez FernándezPilar García Suárez

Lucía Fernández Llamas

Diana Gutiérrez Fernández

Lucía Escobedo Martín.

 

SupervisorsTitlePhD StudentOrganisationDate of VivaClassification
Beatriz Martínez FernándezAna Rodríguez GonzálezMolecular response mechanisms to stress applied to the cell wall in Lactococcus lactisClara Roces RodríguezFaculty of Biology, Universidad de Oviedo. 08/02/2013Cum LaudeEuropean Honours.
Pilar García SuárezAna Rodríguez GonzálezThe potential of phages and phage proteins for eliminating biofilms formed by StaphylococciDiana Gutiérrez Fernández Faculty of Biology, Universidad de Oviedo 8/5/2015DistinctionInternational 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 VázquezInstituto Tecnológico de Veracruz (Veracruz Technological Institute), Mexico 15/06/2015Pass (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
Country: Spain
Organisation: CSIC