Person: Ugarte Ruiz, María
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María
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Ugarte Ruiz
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Universidad Complutense de Madrid
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Item Non-invasive surveillance of shared pathogens in the Eurasian brown bear (Ursus arctos) human interface(One Health, 2024) Herrero García, Gloria; Barroso, Patricia; Dashti, Alejandro; González Barrio, David; Naves, Javier; Fernández Gil, Alberto; Ugarte Ruiz, María; Pérez Sancho, Marta; Royo, Luis José; Carmena, David; De Miguel, Arturo; García Rodríguez, Alberto; Gortázar, Christian; Domínguez Rodríguez, Lucas José; Balseiro, AnaMulti-host communities are perfect scenarios for the emergence and spread of pathogens, threatening the recovery of endangered, isolated, or inbred populations, such as the brown bear (Ursus arctos) in northwestern Spain. The population recovery in recent years has forced bears to occupy highly anthropized areas, increasing their interaction with human and domestic animals, with potential consequences for global health. During 2022–2023 a survey of parasites, bacteria and viruses shared between wildlife, domestic animals and humans was performed in this population using non-invasive surveillance, i.e., bear fecal samples (n = 73) and sponge-based sampling of trees (n = 42; 14 rubbed trees and 28 control trees). Pathogen detection rates were defined as the percentage of qPCR or culture-positive samples. Generalized linear models were fitted to assess their relationship with environmental variables including dispersion of the human population, and percentage of agricultural and periurban habitats in a 6 km-buffer around each sample. Canine Adenovirus type 1 (45.2%), Giardia spp. (15.1%), Salmonella spp. (12.3%), and extended-spectrum-beta-lactamases (ESBL) Escherichia coli (1.4%) were identified in fecal samples. In contrast, only five sponges from three rubbed and two control trees resulted positive to E. coli (14.3%). The results suggest that several pathogens are common in the Cantabrian brown bear population and that anthropization of the territory modulates their prevalence and richness. The effective design of management programs for bear conservation will require a one-health approach, in which genetic analysis of non-invasive samples can be key tools for the sanitary surveillance at the wildlife-livestock-human interface.Item Carbapenemase-Producing Elizabethkingia Meningoseptica from Healthy Pigs Associated with Colistin Use in Spain(Antibiotics, 2019) Miguela Villoldo, Pedro; Hernández, Marta; Moreno Romo, Miguel Ángel; Rodríguez-Lázaro, David; Quesada, Alberto; Ugarte Ruiz, María; Domínguez Rodríguez, Lucas JoséCarbapenems are considered last-resort antimicrobials, especially for treating infections involving multidrug-resistant Gram-negative bacteria. In recent years, extended-spectrum β-lactamase (ESBL) and carbapenemase-producing Gram-negative bacteria have become widespread in hospitals, community settings, and the environment, reducing the range of effective therapeutic alternatives. The use of colistin to treat infection caused by these multi-drug bacteria may favour the selection and persistence of carbapenem-resistant bacteria. In this study, it is described, for the first time to our knowledge, a carbapenemase-producing isolate of Elizabethkingia meningoseptica from healthy pigs in Spain. The isolate we report was recovered during a study to detect colistin-resistant bacteria from faecal samples of healthy food-production animals using a chromogenic selective medium. Unexpectedly, we found an isolate of Elizabethkingia meningoseptica with high Minimum Inhibitory Concentration (MIC) values for several antibiotics tested. Molecular analysis did not show any mcr family genes related with colistin resistance, but two carbapenemase genes, blaB-12_1 and blaGOB-17_1, were detected. This finding in healthy animals could suggest that colistin may favour the selection and persistence of carbapenem-resistant bacteria.- Project number: 265
Item SWI@UCM 2.0: Consolidación de Small World Initiative: descubrimiento y uso racional de antibióticos mediante aprendizaje-servicio en la Comunidad de Madrid(2018) Jimenez Cid, Víctor; Rodriguez Escudero, María Isabel; Díez Orejas, Rosalía María; Molina Martín, María; Rodríguez Fernández, Carmina; Navarro García, Federico; Arroyo, Francisco Javier; Román González, Elvira; Martín Brieva, Humberto; Fernández-Acero Bascones, Teresa; Sanz Santamaría, Ana Belén; Díaz del Toro, Silvia; Calvo de Pablo, Pilar; Patiño Álvarez, Aurora Belén; González Zorn, Bruno; Suárez Rodríguez, Mónica; Goyache Goñi, Joaquín; Escudero García-Calderón, José Antonio; Prieto Prieto, Antonio Daniel; Ugarte Ruiz, María; Gil-Serna, JessicaDurante el curso anterior un equipo de la UCM en el marco de un proyecto previo INNOVA-Docencia UCM instauró en España de manera pionera la iniciativa de aprendizaje activo Small World Initiative, de origen norteamericano. Los objetivos de esta iniciativa son (1) Crear cultura científica y acercar la investigación biomédica a niveles educativos en los que los estudiantes tienen aún capacidad de decisión sobre su futura orientación formativa con el fin de fomentar la vocación en I+D; y (2) Promover la concienciación social sobre el uso racional de los antibióticos y la amenaza de la resistencia bacteriana a estos fármacos. En el entorno español se propuso implementar esta estrategia mediante Aprendizaje-Servicio. En esta segunda edición (SWI@UCM 2.0) se ha trabajado en la consolidación, expansión y mejora del proyecto, con énfasis en la integración de los diversos niveles educativos que integran el proyecto (universitario y preuniversitario) y la divulgación científica. Item Effects on Intestinal Mucosal Morphology, Productive Parameters and Microbiota Composition after Supplementation with Fermented Defatted Alperujo (FDA) in Laying Hens(Antibiotics, 2019) Rebollada Merino, Agustín Miguel; Bárcena Asensio, Carmen; Ugarte Ruiz, María; Porras González, Néstor; Mayoral Alegre, Francisco Jesús; Tomé Sánchez, Irene; Rodríguez Bertos, Antonio Manuel; Domínguez Rodríguez, Lucas JoséThe olive oil sector is currently adapting its traditional function to also become a supplier of high-value by-products that possess antioxidant, anti-inflammatory and antimicrobial properties. In this study, we evaluated the effect of the fermented defatted alperujo (FDA) on the intestinal health of laying hens. The morphology of the duodenal and cecal mucosa, the composition of the intestinal microbiota and the productivity of a batch of laying hens were evaluated after FDA supplementation. At early life stages, significant differences (p < 0.001) were observed in duodenal villi height and in crypt depth of both the duodenum and the cecum in the FDA-supplemented group, indicating improved intestinal health in this group. Microbiota composition in the hatchery group supplemented with FDA had a higher abundance of Actinobacteria, Firmicutes and Proteobacteria, and higher bacterial diversity. During the production period, significant differences (p < 0.05) were observed in the number of broken eggs from the supplemented group. We conclude that FDA supplementation improves the absorption capacity of the intestinal mucosa and modifies the intestinal microbiota to favor a greater immune response, leading to an increase in egg production.Item Detección y Caracterización de "Campylobacter" procedentes de animales, alimentos y agua residual(2015) Ugarte Ruiz, María; Domínguez Rodríguez, Lucas; Porrero Calonge, María Concepción; Wassenaar, TrudyEn el presente trabajo de tesis doctoral se ha estudiado la presencia de Campylobacter termófilos en muestras obtenidas en animales y alimentos en distintas fases de la cadena alimentaria, en efluentes urbanos y en heces de animales de vida libre ubicados en un entorno geográfico compartido. La finalidad fue comparar diferentes protocolos de detección de Campylobacter y obtener información sobre su epidemiología. Adicionalmente, se propuso el estudio de la presencia de un sistema de secreción de proteínas: sistema de secreción tipo VI o SST6, relacionado con la patogenicidad y supervivencia de algunas cepas de Campylobacter. Los resultados obtenidos mediante cultivo, implican la presencia de Campylobacter termófilos en prácticamente la totalidad de las matrices analizadas, con la excepción de la cabra montés. La proporción de positivos también varió en función de los distintos protocolos de detección utilizados, funcionando mejor la siembra directa en muestras de contenido cecal y piel de cuello, y el enriquecimiento en muestras de pechuga de pollo fresca envasada y efluentes urbanos. Por otro lado, la PCR en tiempo real confirmó la presencia de ADN de C. jejuni y/o C. coli en un elevado porcentaje de muestras en todos los estudios. n cuanto a las especies detectadas, la distribución de las mismas varió en función de la matriz analizada, siendo mayor la proporción de aislados de C. coli en muestras de contenido cecal, pechuga de pollo y efluentes urbanos, mientras que en piel de cuello la proporción de C. jejuni fue mayor. Además, la proporción de especies detectadas se vio influenciada significativamente por el protocolo de detección empleado, ya que el enriquecimiento favoreció la detección de C. coli, mientras que la siembra directa obtuvo una mayor proporción de C. jejuni. En el estudio llevado a cabo en animales de vida libre, se detectó C. jejuni en jabalí y ganado vacuno criado en extensivo, aunque C. coli sólo se identificó en muestras de jabalí. También se observó C. lanienae en ambos grupos, siendo la especie predominante en el jabalí. Estos resultados indicarían que podría existir un flujo potencial de Campylobacter entre jabalí y ganado bovino criado en extensivo, aunque la cabra montés no desempeñaría ningún papel en la epidemiología de esta bacteria.Por otro lado, la caracterización genética de los aislados de la cadena alimentaria indica que la matriz con mayor riqueza de genotipos es la piel de cuello. Así, los aislados de Campylobacter encontrados en las muestras de pechuga y piel de cuello podrían proceder tanto del mismo animal y/o lote, como de contaminaciones cruzadas con cepas de animales y/o lotes diferentes en el matadero. Asimismo, la riqueza de genotipos encontrada fue disminuyendo al final de la cadena de producción, viéndose influenciada por el protocolo de cultivo empleado. En lo referente a la caracterización de la resistencia a antimicrobianos en efluentes urbanos, se identificó una mayor proporción de resistencias en aislados de C. coli que en aislados de C. jejuni. El hecho de que los patrones de resistencia detectados sean similares a los descritos en aislados humanos, sugiere la posibilidad de utilizar esta matriz como un buen indicador del nivel de resistencias en cepas de Campylobacter de la comunidad. Por último, se detectaron los trece genes que conforman el SST6 en el 14 por ciento de los aislados de C. jejuni analizados. Se identificaron aislados positivos en todas las matrices de la cadena alimentaria, aunque ninguno de los aislados de efluentes urbanos fue positivo. Los resultados indican que la presencia del gen hcp ¿hemolysin coregulated protein¿, utilizado como indicador de la presencia de SST6, no implica la presencia del sistema completo, ya que algunas cepas poseían dicho gen y carecían del resto de genes del sistema.Item Rapid differentiation of Staphylococcus aureus subspecies based on MALDI-TOF MS profiles(2018) Pérez Sancho, Marta; Vela Alonso, Ana Isabel; Horcajo Iglesias, María Del Pilar; Ugarte Ruiz, María; Domínguez Rodríguez, Lucas José; Fernández-Garayzábal Fernández, José Francisco; Fuente López, Ricardo De LaStaphylococcus aureus encompasses 2 subspecies ( aureus and anaerobius) with significant differences in their epidemiology and pathogenicity. We evaluated the suitability of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for the rapid identification of both subspecies using a panel of 52 S. aureus isolates (30 subsp. anaerobius and 22 subsp. aureus) recovered from different origins, countries, and years. The on-board library identification system correctly identified 42 of 52 (81%) S. aureus isolates at the species level with score values >2.0. Limited performance was observed for differentiation of S. aureus subspecies (particularly subsp. anaerobius). Visual inspection of MALDI-TOF MS profiles identified 5 subspecies-specific mass peaks ( m/ z 3430 and 6861 in S. aureus subsp. anaerobius, and m/ z 4046, 6890, and 8093 in S. aureus subsp. aureus) with 100% sensitivity and specificity values, which is potentially useful for differentiating these subspecies. The suitability of 3 models, Genetic Analysis (GA), Quick Classifier (QC), and Supervised Neural Network, for automatic identification of both subspecies was evaluated using the Recognition Capability (RC) and Cross Validation (CV) values provided by the on-board ClinProTools software. The GA and QC models reached RC and CV values of 100%. Both models were externally validated using a panel of 26 S. aureus isolates of both subspecies, with both models correctly classifying all isolates of both subspecies. MALDI-TOF MS coupled with ClinProTools software represents a rapid and simple approach for S. aureus subspecies discrimination.Item Non-invasive surveillance of shared pathogens in the Eurasian brown bear (Ursus arctos) human interface(One Health, 2024) Herrero García, Gloria; Barroso, Patricia; Dashti, Alejandro; González Barrio, David; Naves, Javier; Fernández-Gil, Alberto; Ugarte Ruiz, María; Pérez Sancho, Marta; Royo, Luis José; Carmena, David; De Miguel, Arturo; García Rodríguez, Alberto; Gortázar, Christian; Domínguez Rodríguez, Lucas José; Balseiro, AnaMulti-host communities are perfect scenarios for the emergence and spread of pathogens, threatening the recovery of endangered, isolated, or inbred populations, such as the brown bear (Ursus arctos) in northwestern Spain. The population recovery in recent years has forced bears to occupy highly anthropized areas, increasing their interaction with human and domestic animals, with potential consequences for global health. During 2022-2023 a survey of parasites, bacteria and viruses shared between wildlife, domestic animals and humans was performed in this population using non-invasive surveillance, i.e., bear fecal samples (n = 73) and sponge-based sampling of trees (n = 42; 14 rubbed trees and 28 control trees). Pathogen detection rates were defined as the percentage of qPCR or culture-positive samples. Generalized linear models were fitted to assess their relationship with environmental variables including dispersion of the human population, and percentage of agricultural and periurban habitats in a 6 km-buffer around each sample. Canine Adenovirus type 1 (45.2%), Giardia spp. (15.1%), Salmonella spp. (12.3%), and extended-spectrum-beta-lactamases (ESBL) Escherichia coli (1.4%) were identified in fecal samples. In contrast, only five sponges from three rubbed and two control trees resulted positive to E. coli (14.3%). The results suggest that several pathogens are common in the Cantabrian brown bear population and that anthropization of the territory modulates their prevalence and richness. The effective design of management programs for bear conservation will require a one-health approach, in which genetic analysis of non-invasive samples can be key tools for the sanitary surveillance at the wildlife-livestock-human interface.Item Colistin Selection of the Mcr-1 Gene in Broiler Chicken Intestinal Microbiota(Antibiotics, 2021) Miguela Villoldo, Pedro; Moreno, Miguel A.; Rebollada Merino, Agustín Miguel; Rodríguez Bertos, Antonio Manuel; Hernández, Marta; Rodríguez Lázaro, David; Gallardo, Alejandro; Quesada, Alberto; Goyache Goñi, Joaquín ; Ugarte Ruiz, María; Domínguez Rodríguez, Lucas JoséColistin has a long story of safe use in animals for the treatment and prevention of certain bacterial diseases. Nevertheless, the first description of the mcr-1 gene showed that colistin resistance can spread by horizontal gene transfer and changed the landscape. This study aimed to assess the effect of colistin administration on the dispersion of resistance in the microbiota of day-old broiler chicks and how the presence of mcr-1 genes influences the spread of colistin resistance determinants. In this study, 100 one-day-old chicks were divided into four groups of 25 animals (G1, G2, G3, and G4). Animals from G3/G4 were challenged with mcr-1-carrying Salmonella (day 7), while colistin (600 mg/L) was administered daily to G2/G4 animals through drinking water (from day 8 to day 15). Two quantitative PCR assays were performed to compare the amount of Salmonella and mcr-1 that were present in the caecal samples. We observed that levels of mcr-1 were higher in G3/G4 animals, especially G4, due to the spread of mcr-1-carrying Salmonella. On day 21, Salmonella levels decreased in G4, reaching similar values as those for G3, but mcr-1 levels remained significantly higher, suggesting that colistin may accelerate the spreading process when mcr-1-carrying bacteria reach the gut.Item Method Comparison for Enhanced Recovery, Isolation and Qualitative Detection of C. jejuni and C. coli from Wastewater Effluent Samples(International Journal of Environmental Research and Public Health, 2015) Ugarte Ruiz, María; Florez Cuadrado, Diego; Wassenaar, Trudy M; Porrero, Maria Concepcion; Domínguez Rodríguez, Lucas JoséSeeking a sensitive protocol, culture-dependent methods were compared to detect thermophilic Campylobacter species in untreated urban effluents. We evaluated various combinations of selective media, with and without an enrichment steps, as well as an extra filtration step. Culture-independent real-time quantitative PCR was also included and all detected isolates underwent antimicrobial susceptibility testing. All tested water samples contained Campylobacter DNA, but only 64% were positive after culture. Although enrichment using Preston broth resulted in better recovery of potentially stressed Campylobacter than Bolton or Campyfood broth (CFB), there was no significant increase in efficiency compared to direct plating. The type of selective agar media used, on the other hand, had a significant effect, with CASA plates performing better than mCCDA or CFA ones. Inclusion of an enrichment step increased the ratio of C. coli vs. C. jejuni being isolated. Resistances against all antimicrobials tested were observed in C. coli, but fewer instances of resistance were found in C. jejuni isolates.Item Cecal Reduction of Brachyspira and Lesion Severity in Laying Hens Supplemented with Fermented Defatted ‘Alperujo’(Fermentation, 2022) Rebollada Merino, Agustín Miguel; Ugarte Ruiz, María; Alberto Gómez-Buendía; Bárcena Asensio, María Carmen; García Benzaquén, Nerea; Domínguez Rodríguez, Lucas José; Rodríguez Bertos, Antonio ManuelAntimicrobial resistance demands the development of therapeutic alternatives such as prebiotics, probiotics, and nutraceuticals. The aim of this study was to assess the antimicrobial proprieties of the nutraceutical fermented defatted “alperujo”, derived from olive oil production, in a laying hen farm (n = 122,250) endemic with avian intestinal spirochetosis (Brachyspira spp.). Part of the batch (n = 1440) was divided into six groups of 240 hens each that included 80 or 108-week-old laying hens, supplemented with 0%, 2%, or 6% fermented defatted ‘alperujo’ for a month. At the end of the experiment, eight hens from each group were autopsied and cecal content was subjected to (i) Brachyspira culture and species identification by PCRs, and (ii) direct DNA extraction and Brachyspira qPCR. Furthermore, the ceca were processed for histopathology. Microbiological isolation revealed B. pilosicoli and B. hyodysenteriae co-infection in all groups. The 80-week-old hen group 2% supplemented showed a reduction in the cecal Brachyspira content (qPCR) compared with non-supplemented hens. Cecal histopathology showed a diffuse mild infiltration of lymphocytes, plasma cells, and heterophils; and hyperplasia of the gut-associated lymphoid tissue hyperplasia which decreased in severity in 80-week-old supplemented hens. The reduction in Brachyspira colonization and the severity of the lesions observed in supplemented hens highlights a potential protective function against avian intestinal spirochetosis.