Influence of Sodium Chloride on the Behaviour of Pseudomonas fluorescens in Ripened Sheep Cheese Simone Lopes, Manuela Vida, Cláudia Correia, Jaime Fernandes, Sandra Gomes, Ana Fernando, Rafael Tabla, Nuno Alvarenga Microorganisms, 2025 Ewe’s milk cheese produced from raw milk holds cultural and economic importance in Southern European countries; however, it poses microbiological challenges. Among spoilage microorganisms, Pseudomonas fluorescens is particularly concerning due to thermostable enzymes that impair the texture, aroma, and stability of cheese, even under refrigeration and salinity. This study evaluated the influence of sodium chloride concentration on Pseudomonas fluorescens given the pivotal role of salt in ensuring cheese stability and safety. Cheeses inoculated with Pseudomonas fluorescens were produced under an experimental design that combined three ripening temperatures with four salt concentrations. Physicochemical composition and microbiological stability were assessed at the end of ripening (20 days). Results showed that the ripening temperature emerged as the most determinant factor, influencing microbial viability and increasing solid retention, proteolysis, and dehydration, leading to harder cheeses. Low temperatures without salt favoured surface colour defects, whereas, although high salt levels contributed to partial control of Pseudomonas spp., they also delayed ripening, resulting in cheeses with a pale, uncharacteristic appearance. Conversely, moderate salinity (2%) combined with higher ripening temperatures promoted uniform maturation, resulting in a stable texture and appearance free of defects. These findings highlight the need to balance salt and ripening conditions to optimise quality and safety in traditional raw ewe’s milk cheeses.
ECOPHAGE: Combating Antimicrobial Resistance Using Bacteriophages for Eco-Sustainable Agriculture and Food Systems Pilar García, Rafael Tabla, Hany Anany, Roberto Bastias, Lone Brøndsted, Susana Casado, Pablo Cifuentes, John Deaton, Thomas G. Denes, Mohammad Aminul Islam, Rob Lavigne, Andrea I. Moreno-Switt, Natsuko Nakayama, Cristina Muñoz Madero, Alexander Sulakvelidze, Antonet M. Svircev, Jeroen Wagemans, Elena G. Biosca, Dácil Rivera Viruses, 2023 The focus of this meeting was to discuss the suitability of using bacteriophages as alternative antimicrobials in the agrifood sector. Following a One Health approach, the workshop explored the possibilities of implementing phage application strategies in the agriculture, animal husbandry, aquaculture, and food production sectors. Therefore, the meeting had gathered phage researchers, representatives of the agrifood industry, and policymakers to debate the advantages and potential shortcomings of using bacteriophages as alternatives to traditional antimicrobials and chemical pesticides. Industry delegates showed the latest objectives and demands from consumers. Representatives of regulatory agencies (European Medicines Agency (EMA) and Spanish Agency of Medicines and Health Products (AEMPS)) presented an update of new regulatory aspects that will impact and support the approval and implementation of phage application strategies across the different sectors.
Use of gaseous ozone in soft cheese ripening: Effect on the rind microorganisms and the sensorial quality Rafael Tabla, Isidro Roa Lwt, 2022 The uncontrolled growth of potential spoilage microorganisms in cheese surfaces can deteriorate cheese quality, especially filamentous fungi. This study aimed to evaluate the effect of several gaseous ozone treatments during the ripening of soft cheese on the growth of cheese rind microorganisms at different production scales. On a pilot plant scale, the cheeses were inoculated with potential spoilage microorganisms (Mucor plumbeus, Kluyveromyces marxianus, and Pseudomonas fluorescens), while on an industrial scale, naturally present microorganisms were evaluated. Ozone treatments showed a considerable fungistatic effect on cheese surface molds at both pilot and industrial scale. However, the effect of ozone on yeast growth was only observed at pilot plant scale, while Pseudomonas ssp. was not affected by the ozone concentrations used in this study. The sensory quality of the cheese was also assessed. Cheese typicality was not affected by gaseous ozone. However, the ozonized samples had significantly higher scores than the control samples for rind appearance and color. Hence, ozonation can be considered an alternative approach to control mold growth in soft cheese during ripening and prevent cheese rind discoloration.
Effectiveness of a bacteriophage cocktail in reducing cheese early blowing caused by Escherichia coli Rafael Tabla, Antonia Gómez, José Emilio Rebollo, Felipe Molina, Isidro Roa Lwt, 2022 Gas production and subsequent food spoilage from the appearance of fissures, cracks or holes are undesirable consequences of saprophytic E. coli growth in cheese. Hence, the development of E. coli growth inhibitors is of special interest in soft and raw milk cheese production. This study investigates the inhibitory effect of a cocktail of four selected coliphages on the growth of E. coli in cheese and its ability to inhibit cheese early blowing. The microbiological analysis showed that the evolution of total bacterial counts during cheese-making and ripening was comparable among control and phage inoculated cheeses. Nevertheless these results also revealed a reduction in E. coli growth at the beginning of fermentation with phage treatment. Additionally, image analysis of cheese slices showed a noteworthy reduction in the number of cheese eyes and the area occupied by gas holes throughout cheese ripening. Thus, the use of bacteriophages as biocontrol agents provides an effective approach to combat cheese early blowing without noticeably altering the microbiological or physicochemical parameters of these fermented foods.
A New Pipeline for Designing Phage Cocktails Based on Phage-Bacteria Infection Networks Felipe Molina, Alfredo Simancas, Manuel Ramírez, Rafael Tabla, Isidro Roa, José Emilio Rebollo Frontiers in Microbiology, 2021 In recent years, the spread of antibiotic-resistant bacteria and efforts to preserve food microbiota have induced renewed interest in phage therapy. Phage cocktails, instead of a single phage, are commonly used as antibacterial agents since the hosts are unlikely to become resistant to several phages simultaneously. While the spectrum of activity might increase with cocktail complexity, excessive phages could produce side effects, such as the horizontal transfer of genes that augment the fitness of host strains, dysbiosis or high manufacturing costs. Therefore, cocktail formulation represents a compromise between achieving substantial reduction in the bacterial loads and restricting its complexity. Despite the abovementioned points, the observed bacterial load reduction does not increase significantly with the size of phage cocktails, indicating the requirement for a systematic approach to their design. In this work, the information provided by host range matrices was analyzed after building phage-bacteria infection networks (PBINs). To this end, we conducted a meta-analysis of 35 host range matrices, including recently published studies and new datasets comprising Escherichia coli strains isolated during ripening of artisanal raw milk cheese and virulent coliphages from ewes’ feces. The nestedness temperature, which reflects the host range hierarchy of the phages, was determined from bipartite host range matrices using heuristic (Nestedness Temperature Calculator) and genetic (BinMatNest) algorithms. The latter optimizes matrix packing, leading to lower temperatures, i.e., it simplifies the identification of the phages with the broadest host range. The structure of infection networks suggests that generalist phages (and not specialist phages) tend to succeed in infecting less susceptible bacteria. A new metric (Φ), which considers some properties of the host range matrices (fill, temperature, and number of bacteria), is proposed as an estimator of phage cocktail size. To identify the best candidates, agglomerative hierarchical clustering using Ward’s method was implemented. Finally, a cocktail was formulated for the biocontrol of cheese-isolated E. coli, reducing bacterial counts by five orders of magnitude.
Diversity and Local Coadaptation of Escherichia coli and Coliphages From Small Ruminants Felipe Molina, Alfredo Simancas, Rafael Tabla, Antonia Gómez, Isidro Roa, José Emilio Rebollo Frontiers in Microbiology, 2020 Bacteriophages are highly specific predators that drive bacterial diversity through coevolution while striking tradeoffs among preserving host populations for long-term exploitation and increasing their virulence, structural stability, or host range. Escherichia coli and other coliform bacteria present in the microbiota of milk and during early ripening of raw milk cheeses have been linked to the production of gas, manifested by the appearance of eyes, and the development of off-flavors; thus, they might cause early blowing and cheese spoilage. Here, we report the characterization of coliphages isolated from manure from small ruminant farms and E. coli strains isolated from goat and sheep raw milk cheese. Additionally, the virulence and host range of locally isolated and laboratory collection phages were determined by comparing the susceptibility of E. coli strains from different sources. In agreement with the high genetic diversity found within the species E. coli, clustering analysis of whole-cell protein revealed a total of 13 distinct profiles but none of the raw milk cheese isolates showed inhibition of growth by reference or water-isolated coliphages. Conversely, 10 newly isolated phages had a broad host range (i.e., able to lyse ≥50% of bacterial hosts tested), thus exhibiting utility for biocontrol and only one cheese-isolated E. coli strain was resistant to all the phages. Whereas there was a high positive correlation between bacterial susceptibility range and lysis intensity, the phages virulence decreased as range increased until reaching a plateau. These results suggest local gene-for-gene coevolution between hosts and phages with selective tradeoffs for both resistance and competitive ability of the bacteria and host-range extension and virulence of the phage populations. Hence, different phage cocktail formulations might be required when devising long-term and short-term biocontrol strategies.
Optimising detection of acidification kinetics diversity in Lactococcus lactis subsp. lactis using SDS-PAGE protein fingerprinting as screening method Rafael Tabla, Felipe Molina, José E. Rebollo, Isidro Roa International Journal of Dairy Technology, 2019 Cell‐free extracts from 157 Lactococcus lactis strains isolated from Artisan cheese were screened by protein fingerprinting previous to their technological characterisation. The strains were classified according to their electrophoretic patterns into five groups. A set of strains representing the different clusters were selected to study their acidifying activity in milk. Time and rate feature points, as well as the shape of the acidification curves, resulted in six different fermentation kinetics, mostly consistent with the electrophoretic groups. Thus, selection of native strains as starter cultures based on their acidifying activity could be optimised by protein fingerprinting.
Early blowing in raw goats' milk cheese: Gas production capacity of Enterobacteriaceae species present during manufacturing and ripening Rafael Tabla, Antonia Gómez, Alfredo Simancas, José Emilio Rebollo, Felipe Molina, Isidro Roa Journal of Dairy Research, 2018 The aim of this study was to identify the main Enterobacteriaceae species responsible for early gas blowing during curdling and the first week of ripening in raw goats’ milk cheese. Two batches of raw goats’ milk cheese were selected. One of them showed early blowing within the first 24 h of cheese ripening while the other showed no alteration. Although initial levels of Enterobacteriaceae were similar in defective and non-defective cheese, their dynamics (growth and disappearance rates of the species detected) were different. Klebsiella oxytoca and Enterobacter cloacae were the main species in the defective curd, whereas Buttiauxela spp. was predominant in normal curd. Hafnia alvei was the prevailing isolated species for both normal and defective cheese throughout the ripening process. The highest gas production was rendered by K. oxytoca and H. alvei, mainly isolated from curd and cheese. However, other species relevant in milk or curd, like Pantoea ssp. or Buttiauxela spp. were considered as low gas producers. The analysis of digitalized images of cheese showed that most of the cheese eyes were formed before the first week of ripening, although this process continued during maturation.According to the species found in the defective and non-defective cheese, their proportions at different ripening stages, their ability to produce gas and eye formation, K. oxytoca might be considered the most likely responsible for early blowing in raw goats’ milk cheeses; while H. alvei increased the eyes number in the later stages of the ripening period.
From rind to core: metagenomic and response surface insights into how salt and ripening temperature shape the microbial ecology of ewe’s milk cheese S Lopes, M Vida, C Correia, J Fernandes, S Gomes, A Fernando, R Tabla, ... International Dairy Journal, 106556 , 2026 2026
Influence of Sodium Chloride on the Behaviour of Pseudomonas fluorescens in Ripened Sheep Cheese S Lopes, M Vida, C Correia, J Fernandes, S Gomes, A Fernando, R Tabla, ... Microorganisms 13 (12), 2693 , 2025 2025 Citations: 1
Pseudomonas en corteza de quesos de pasta blanda: dentificación y producción de pigmentos JR Pinilla, Á Acedo, R Tabla XLVII Jornadas de viticultura y enología de la tierra de barros. VII … , 2025 2025
Ecophage: combating antimicrobial resistance using bacteriophages for eco-sustainable agriculture and food systems P García, R Tabla, H Anany, R Bastias, L Brøndsted, S Casado, ... Viruses 15 (11), 2224 , 2023 2023 Citations: 15
Proteólisis en Quesos de La Serena DOP: efecto del formato y estacionalidad T Montero, R Tabla XLIV Jornadas de viticultura y enología de la Tierra de Barros; IV Congreso … , 2023 2023
Use of gaseous ozone in soft cheese ripening: Effect on the rind microorganisms and the sensorial quality R Tabla, I Roa Lwt 170, 114066 , 2022 2022 Citations: 20
Effectiveness of a bacteriophage cocktail in reducing cheese early blowing caused by Escherichia coli R Tabla, A Gomez, JE Rebollo, F Molina, I Roa LWT 153, 112430 , 2022 2022 Citations: 18
A new pipeline for designing phage cocktails based on phage-bacteria infection networks F Molina, A Simancas, M Ramírez, R Tabla, I Roa, JE Rebollo Frontiers in microbiology 12, 564532 , 2021 2021 Citations: 68
A new pipeline for designing phage cocktails based on phage-bacteria infection networks FR Molina Rodríguez, A Simancas, M Ramírez Fernández, R Tabla, I Roa, ... 2021
Corrigendum: Diversity and Local Coadaptation of Escherichia coli and Coliphages From Small Ruminants F Molina, A Simancas, R Tabla, A Gómez, I Roa, JE Rebollo Frontiers in Microbiology 11, 623135 , 2020 2020 Citations: 1
Diversity and Local Coadaptation of Escherichia coli and Coliphages From Small Ruminants F Molina, A Simancas, R Tabla, A Gómez, I Roa, JE Rebollo Frontiers in Microbiology 11, 564522 , 2020 2020 Citations: 16
Cynara cardunculus as a Multiuse Crop G Mauromicale, GR Pesce, MD Curt, J Fernández, J González, ... The Globe Artichoke Genome, 65-98 , 2019 2019 Citations: 23
Optimising detection of acidification kinetics diversity in Lactococcus lactis subsp. lactis using SDS‐PAGE protein fingerprinting as screening method R Tabla, F Molina, JE Rebollo, I Roa International Journal of Dairy Technology 72 (2), 287-294 , 2019 2019 Citations: 2
Fabrico de queijos de ovelha de pasta mole a partir de leite pasteurizado e de coalho vegetal R Tabla Sevillano, T Montero Muñoz, M Garrido Álvarez, M López Cedó, ... 2019
Queijos de ovelha de pasta mole fabricados a partir de leite pasteurizado e de coalho vegetal CM Pintado, AM Rodrigues, T Montero Muñoz, M Garrido Álvarez, ... 2019
Early blowing in raw goats’ milk cheese: gas production capacity of Enterobacteriaceae species present during manufacturing and ripening R Tabla, A Gómez, A Simancas, JE Rebollo, F Molina, I Roa Journal of Dairy Research 85 (3), 331-338 , 2018 2018 Citations: 20
Biological surface active compounds in dairy industry: preliminary analysis of an emulsifier protein A Simancas, F Molina, M Murillo, N Del-Valle, R Tabla, I Roa Exploring Microorganisms: Recent Advances in Applied Microbiology, 125 , 2018 2018 Citations: 1
Isolation and host-specificity characterization of coliphages suitable for biocontrol in dairy industries F Molina, A Simancas, M Díaz, R Nuñez, JE Rebollo, R Tabla, I Roa Explor. Microorg. Recent Adv. Appl. Microbiol 91, 91-95 , 2018 2018 Citations: 2
Enterobacteriaceae species during manufacturing and ripening of semi–hard and soft raw ewe’s milk cheese: Gas production capacity R Tabla, A Gómez, A Simancas, JE Rebollo, F Molina, I Roa Small Ruminant Research 145, 123-129 , 2016 2016 Citations: 56
YaiO, a new target for highly specific detection of Escherichia coli by PCR amplification A Simancas, F Molina, R Tabla, I Roa, JE Rebollo Microbes in the spotlight: Recent progress in the understanding of … , 2016 2016 Citations: 2
MOST CITED SCHOLAR PUBLICATIONS
Improved detection of Escherichia coli and coliform bacteria by multiplex PCR F Molina, E López-Acedo, R Tabla, I Roa, A Gómez, JE Rebollo BMC biotechnology 15 (1), 48 , 2015 2015 Citations: 236
Circadian rhythm of melatonin, corticosterone and phagocytosis: effect of stress C Barriga, MI Martín, R Tabla, E Ortega, AB Rodríguez Journal of pineal research 30 (3), 180-187 , 2001 2001 Citations: 186
A new pipeline for designing phage cocktails based on phage-bacteria infection networks F Molina, A Simancas, M Ramírez, R Tabla, I Roa, JE Rebollo Frontiers in microbiology 12, 564532 , 2021 2021 Citations: 68
Bacteriophage performance against Staphylococcus aureus in milk is improved by high hydrostatic pressure treatments R Tabla, B Martínez, JE Rebollo, J González, MR Ramírez, I Roa, ... International Journal of Food Microbiology 156 (3), 209-213 , 2012 2012 Citations: 68
Effect of high‐pressure processing and thermal treatment on quality attributes and nutritional compounds of “Songold” plum purée F González‐Cebrino, J García‐Parra, R Contador, R Tabla, R Ramírez Journal of Food Science 77 (8), C866-C873 , 2012 2012 Citations: 66
Enterobacteriaceae species during manufacturing and ripening of semi–hard and soft raw ewe’s milk cheese: Gas production capacity R Tabla, A Gómez, A Simancas, JE Rebollo, F Molina, I Roa Small Ruminant Research 145, 123-129 , 2016 2016 Citations: 56
Ibores goat's milk cheese: Microbiological and physicochemical changes throughout ripening M Mas, R Tabla, J Moriche, I Roa, J Gonzalez, JE Rebollo, P Cáceres Le Lait 82 (5), 579-587 , 2002 2002 Citations: 54
Autochthonous starter effect on the microbiological, physicochemical and sensorial characteristics of Ibores goat's milk cheeses J Gonzalez, M Mas, R Tabla, J Moriche, I Roa, JE Rebollo, P Cáceres Le Lait 83 (3), 193-202 , 2003 2003 Citations: 33
Cynara cardunculus as a Multiuse Crop G Mauromicale, GR Pesce, MD Curt, J Fernández, J González, ... The Globe Artichoke Genome, 65-98 , 2019 2019 Citations: 23
Use of gaseous ozone in soft cheese ripening: Effect on the rind microorganisms and the sensorial quality R Tabla, I Roa Lwt 170, 114066 , 2022 2022 Citations: 20
Early blowing in raw goats’ milk cheese: gas production capacity of Enterobacteriaceae species present during manufacturing and ripening R Tabla, A Gómez, A Simancas, JE Rebollo, F Molina, I Roa Journal of Dairy Research 85 (3), 331-338 , 2018 2018 Citations: 20
Effectiveness of a bacteriophage cocktail in reducing cheese early blowing caused by Escherichia coli R Tabla, A Gomez, JE Rebollo, F Molina, I Roa LWT 153, 112430 , 2022 2022 Citations: 18
Diversity and Local Coadaptation of Escherichia coli and Coliphages From Small Ruminants F Molina, A Simancas, R Tabla, A Gómez, I Roa, JE Rebollo Frontiers in Microbiology 11, 564522 , 2020 2020 Citations: 16
Ecophage: combating antimicrobial resistance using bacteriophages for eco-sustainable agriculture and food systems P García, R Tabla, H Anany, R Bastias, L Brøndsted, S Casado, ... Viruses 15 (11), 2224 , 2023 2023 Citations: 15
Microbiological and lipolytic changes in high-pressure-treated raw milk cheeses during refrigerated storage J Rodríguez-Pinilla, G Márquez, R Tabla, R Ramírez, FJ Delgado Dairy Science & Technology 95 (4), 425-436 , 2015 2015 Citations: 15
Salt influence on surface microorganisms and ripening of soft ewe cheese R Tabla, A Gómez, JE Rebollo, I Roa Journal of Dairy Research 82 (2), 215-221 , 2015 2015 Citations: 15
Optimising detection of acidification kinetics diversity in Lactococcus lactis subsp. lactis using SDS‐PAGE protein fingerprinting as screening method R Tabla, F Molina, JE Rebollo, I Roa International Journal of Dairy Technology 72 (2), 287-294 , 2019 2019 Citations: 2
Isolation and host-specificity characterization of coliphages suitable for biocontrol in dairy industries F Molina, A Simancas, M Díaz, R Nuñez, JE Rebollo, R Tabla, I Roa Explor. Microorg. Recent Adv. Appl. Microbiol 91, 91-95 , 2018 2018 Citations: 2
YaiO, a new target for highly specific detection of Escherichia coli by PCR amplification A Simancas, F Molina, R Tabla, I Roa, JE Rebollo Microbes in the spotlight: Recent progress in the understanding of … , 2016 2016 Citations: 2
Influence of Sodium Chloride on the Behaviour of Pseudomonas fluorescens in Ripened Sheep Cheese S Lopes, M Vida, C Correia, J Fernandes, S Gomes, A Fernando, R Tabla, ... Microorganisms 13 (12), 2693 , 2025 2025 Citations: 1
