Search

Este artículo se encuentra disponible en la siguiente URL: https://onlinelibrary.wiley.com/doi/epdf/10.1111/tbed.13737 En este artículo de investigación también participan: Marta Cerdà Cuéllar, Teresa Ayats, Jaume Jordá, Clara Marín y Santiago Vega. This is the pre-peer reviewed version of the following article: Sevilla-Navarro, S., Cerdà-Cuéllar, M., Ayats, T., Jordá, J., Marin, C., & Vega, S. (2021). Characterization of "Salmonella Frintrop" isolated from dromedary camels (Camelus dromedarius). Transboundary and Emerging Diseases, vol. 68, i. 2 (mar.), pp. 742?746, which has been published in final form at https://doi.org/10.1111/tbed.13737. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions. Este es el pre-print del siguiente artículo: Sevilla-Navarro, S., Cerdà-Cuéllar, M., Ayats, T., Jordá, J., Marin, C., & Vega, S. (2021). Characterization of "Salmonella Frintrop" isolated from dromedary camels (Camelus dromedarius). Transboundary and Emerging Diseases, vol. 68, i. 2 (mar.), pp. 742?746, que se ha publicado de forma definitiva en https://doi.org/10.1111/tbed.13737. Different studies have reported the prevalence and antibiotic resistance of Salmonella in dromedaries’ camels and its role in camelid-associated salmonellosis in humans, but little is known about the epidemiology of Campylobacter in dromedaries. Here we investigate the prevalence, genetic diversity and antibiotic resistance of Campylobacter and Salmonella in dromedary camels (Camelus dromedarius). A total of 54 individuals were sampled from two unique dromedary farms located in Tenerife (Canary Islands, Spain). Whilst all the samples were Campylobacter-negative, Salmonella prevalence was 5.5% (3/54) and the only serovar isolated was S. Frintrop. The pulsed field gel electrophoresis analysis revealed a low genetic diversity, with all isolates showing a nearly identical pulsotype (similarity > 95%). Our results indicate that dromedaries’ camels could not be a risk factor for Campylobacter human infection, but seems to be a reservoir for Salmonella transmission. Since camel ride has become one of the main touristic attractions in several countries and its popularity has considerably risen in the last years, a mandatory control, especially for zoonotic pathogens, such as Campylobacter and Salmonella should be implemented. Preprint

The emergence of new pathogens is a major threat to public and veterinary health. Changes in bacterial habitat such as a switch in host or disease tropism are typically accompanied by genetic diversification. Staphylococcus aureus is a multi-host bacterial species associated with human and livestock infections. A microaerophilic subspecies, Staphylococcus aureus subsp. anaerobius, is responsible for Morel’s disease, a lymphadenitis restricted to sheep and goats. However, the evolutionary history of S. aureus subsp. anaerobius and its relatedness to S. aureus are unknown. Population genomic analyses of clinical S. aureus subsp. anaerobius isolates revealed a highly conserved clone that descended from a S. aureus progenitor about 1000 years ago before differentiating into distinct lineages that contain African and European isolates. S. aureus subsp. anaerobius has undergone limited clonal expansion, with a restricted population size, and an evolutionary rate 10-fold slower than S. aureus. The transition to its current restricted ecological niche involved acquisition of a pathogenicity island encoding a ruminant host-specific effector of abscess formation, large chromosomal re-arrangements, and the accumulation of at least 205 pseudogenes, resulting in a highly fastidious metabolism. Importantly, expansion of ~87 insertion sequences (IS) located largely in intergenic regions provided distinct mechanisms for the control of expression of flanking genes, including a novel mechanism associated with IS-mediated anti-anti-sense decoupling of ancestral gene repression. Our findings reveal the remarkable evolutionary trajectory of a host-restricted bacterial pathogen that resulted from extensive remodelling of the S. aureus genome through an array of diverse mechanisms in parallel., Author summary The emergence of new pathogens is a major threat to public and veterinary health. Some bacteria such as Staphylococcus aureus, have the capacity to infect many different host species including humans and livestock while others such as the closely-related S. aureus subsp. anaerobius, associated with a single type of pathology called Morel’s disease in small ruminants, are highly niche-restricted. However, our understanding of the genetic basis for such differences in bacterial host-tropism is very limited. Here, we discovered that S. aureus subsp. anaerobius evolved from an S. aureus ancestor and underwent an array of extensive changes to its genome that accompanied the transition to its current restricted lifestyle. We observed genome decay involving loss of function of hundreds of genes, large intra-chromosomal rearrangements affecting most of the genome, acquisition of a pathogenicity island, and expansion of large numbers of insertion sequences that are inserted at intergenic sites around the genome. Importantly, we found that IS elements affect the expression of neighbouring genes in different ways including a novel mechanism of IS-enabled disruption of ancestral gene repression. Taken together, we provide a remarkable example of radical genomic changes associated with evolutionary transition from a multi-host to highly restricted host ecology.

Conjugation has classically been considered the main mechanism driving plasmid transfer in nature. Yet bacteria frequently carry so-called non-transmissible plasmids, raising questions about how these plasmids spread. Interestingly, the size of many mobilisable and non-transmissible plasmids coincides with the average size of phages (~40 kb) or that of a family of pathogenicity islands, the phage-inducible chromosomal islands (PICIs, ~11 kb). Here, we show that phages and PICIs from Staphylococcus aureus can mediate intra- and inter-species plasmid transfer via generalised transduction, potentially contributing to non-transmissible plasmid spread in nature. Further, staphylococcal PICIs enhance plasmid packaging efficiency, and phages and PICIs exert selective pressures on plasmids via the physical capacity of their capsids, explaining the bimodal size distribution observed for non-conjugative plasmids. Our results highlight that transducing agents (phages, PICIs) have important roles in bacterial plasmid evolution and, potentially, in antimicrobial resistance transmission., Nature Communications, 12, ISSN:2041-1723

Este artículo se encuentra disponible en la siguiente URL: https://academic.oup.com/jid/advance-article/doi/10.1093/infdis/jiaa367/5862450 En esta investigación también participan: Nathalie Aulner, Anne Danckaert, Alain Charbit, Jost Enninga, Pascale Cossart y Javier Pizarro Cerdá. The bacterial pathogen Listeria monocytogenes invades host cells, ruptures the internalization vacuole, and reaches the cytosol for replication. A high-content small interfering RNA (siRNA) microscopy screen allowed us to identify epithelial cell factors involved in L. monocytogenes vacuolar rupture, including the serine/threonine kinase Taok2. Kinase activity inhibition using a specific drug validated a role for Taok2 in favoring L. monocytogenes cytoplasmic access. Furthermore, we showed that Taok2 recruitment to L. monocytogenes vacuoles requires the presence of pore-forming toxin listeriolysin O. Overall, our study identified the first set of host factors modulating L. monocytogenes vacuolar rupture and cytoplasmic access in epithelial cells.

El presente estudio evaluó la actividad antimicrobiana de diferentes aceites esenciales (limón, naranja, mandarina, laurel, eucalipto y burro) en bacterias benéficas como Lactobacillus plantarum ES147 y ATCC 8014, patógenas como Escherichia coli ATCC 25922 y una bacteria alterante de alimentos, Leuconostoc mesenteroides MS1. En los aceites que se registraron halos de inhibición significativo (al menos 10 mm de halo en 3 cepas analizadas) se procedió a determinar las concentraciones mínimas inhibitoria y bactericida. Los aceites esenciales de limón, eucalipto y burro mostraron un efecto bactericida en todas las cepas analizadas. Los aceites esenciales de naranja y mandarina presentaron muy bajo o nulo poder antimicrobiano en las cepas en estudio. Los aceites esenciales de burro, laurel, mandarina y naranja no inhibieron o inhibieron muy poco a las bacterias benéficas y generaron grandes halos de inhibición en bacterias patógenas o alterantes de alimentos. Dichos aceites pueden considerarse para su adición en films o recubrimientos de alimentos. The present study evaluated the antimicrobial activity of different essential oils (lemon, orange, mandarin, laurel, eucalyptus and burro) in beneficial bacteria such as Lactobacillus plantarum ES147 and ATCC 8014, pathogens such as Escherichia coli and a food-altering bacterium, Leuconostoc mesenteroides MS1. Subsequently, the minimal inhibitory concentrations and minimal bactericidal concentrations of the essential oils that had a significant bactericidal effect (at least 10 mm inhibition zone in tree strains) were determined. Lemon, eucalyptus and burro essential oils showed a bactericidal effect in all analyzed strains. Mandarin and orange essential oils presented very low or null antimicrobial inhibition in the strains under study. Laurel and burro essential oils generated significantly lower inhibition halos in beneficial bacteria compared to pathogenic or food-altering bacteria. Such essential oils can be considered for addition in films or food coatings. Fil: Vignola, María Belén. Universidad Tecnológica Nacional. Facultad Reg.san Francisco. Departamento de Ingeniería Química; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina Fil: Serra, Mónica Andrea. Universidad Tecnológica Nacional. Facultad Reg.san Francisco. Departamento de Ingeniería Química; Argentina Fil: Andreatta, Alfonsina Ester. Universidad Tecnológica Nacional. Facultad Reg.san Francisco. Departamento de Ingeniería Química; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina

The abuse and misuse of antibiotics, such as through self-medication, treatment dropout, or cattle fattening, result in the emergence of resistant or multi-resistant pathogenic bacteria. This emergence could be due to an increased selective pressure that kills sensitive strains, new antibiotic resistances appearing at a faster rate than in nature, with higher minimum inhibitory concentrations (MICs), and than the drug discovery process. Awareness and a drop in consumption seem essential to slow down this rate of increased resistance. In this study, we asked ourselves if controlling the use of antibiotics and, hence, a drop in selective pressure are sufficient for resistant pathogenic bacteria to revert to their sensitive condition. To answer this question, we carried out a bibliographic review. The analyzed data suggests that bacteria will compensate for the fitness cost due to their high adaptive capacity. Therefore, bacterial resistance will not be reverted just by lowering the selective pressure. We suggest further research on bacterial evolution at the molecular level is necessary in order to find a solution to this problem as soon as possible. El abuso y el mal uso de los antibióticos como, por ejemplo, la automedicación, el abandono de tratamientos o su utilización para el engorde de animales de granja, da como resultado la aparición de bacterias patógenas resistentes o multirresistentes a antibióticos. Posiblemente, esto se deba a un exceso de presión selectiva que permite eliminar la competencia de cepas sensibles, apareciendo nuevas resistencias a antibióticos a una velocidad mayor que en la naturaleza, con concentraciones mínimas inhibitorias (CMI) más altas y en menos tiempo que el requerido para crear nuevos fármacos. Para frenar este avance, la sensibilización, la concienciación y la bajada en el consumo parecen fundamentales. En este estudio, nos preguntamos si el control del uso de los antibióticos y, por tanto, la bajada en la presión selectiva es suficiente para que las bacterias patógenas resistentes o multirresistentes reviertan a su condición de sensibles. Para dar respuesta a esta pregunta, hemos llevado a cabo un análisis bibliográfico. Tras analizar los datos, todo parece indicar que las bacterias resistentes no van a revertir al estado de sensibles tras bajar solamente la presión selectiva debido a su gran capacidad de adaptación. Sugerimos, entre otras cosas, una mayor investigación sobre la evolución bacteriana a nivel molecular, para encontrar cuanto antes una solución a este problema. O abuso e uso indevido de antibióticos, como por exemplo, a automedicação, o abandono de tratamentos ou o uso para engordar animais de criação, têm como resultado o aparecimento de bactérias patogénicas resistentes ou multirresistentes aos antibióticos. Provavelmente, devido ao excesso de pressão seletiva que permite eliminar a competência de cepas sensíveis, aparecendo novas resistências aos antibióticos com uma rapidez superior à que ocorre na natureza, com concentrações mínimas inibitórias (CIM) mais altas e em menos tempo do que o necessário, para a criação de novos medicamentos. Para conter este avanço é fundamental, a sensibilização, a conscientização e a diminuição do consumo. Neste estudo, questionamos se o controle do uso de antibióticos e portanto, a redução da pressão seletiva é suficiente, para que as bactérias patogénicas resistentes ou multirresistentes voltem à sua condição de sensíveis. Para responder a esta pergunta, realizamos uma análise bibliográfica. Depois de analisar os dados, tudo parece indicar que depois de baixar apenas a pressão seletiva, as bactérias resistentes não voltarão ao seu estado sensível, devido à sua grande adaptabilidade. Sugerimos, entre outras coisas, estudos adicionais sobre a evolução bacteriana a nível molecular, para encontrar uma solução para este problema, o mais breve possível.

Este artículo se encuentra disponible en la siguiente URL: https://onlinelibrary.wiley.com/doi/10.1111/tbed.13470 This is the peer reviewed version of the following article: Sevilla, E., Marín, C., Delgado-Blas, JF., González-Zorn, B., Vega, S., Kuijper, E., Bolea, R. & Mainar-Jaime, RC. Wild griffon vultures (Gyps fulvus) fed at supplementary feeding stations: Potential carriers of pig pathogens and pig-derived antimicrobial resistance?. Transbounddary and Emergency Diseases, vol. 67, i. 3 (may 2020), pp. 1295-1305, which has been published in final form at https://doi.org/10.1111/tbed.13470. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions. Este es el post-print del siguiente artículo: Sevilla, E., Marín, C., Delgado-Blas, JF., González-Zorn, B., Vega, S., Kuijper, E., Bolea, R. & Mainar-Jaime, RC. Wild griffon vultures (Gyps fulvus) fed at supplementary feeding stations: Potential carriers of pig pathogens and pig-derived antimicrobial resistance?. Transbounddary and Emergency Diseases, vol. 67, i. 3 (may 2020), pp. 1295-1305, que se ha publicado de forma definitiva en https://doi.org/10.1111/tbed.13470 The carriage of two important pathogens of pigs, i.e. enterotoxigenic E. coli (ETEC) and Clostridioides difficile, was investigated in 104 cloacal samples from wild griffon vultures (Gyps fulvus) fed on pig carcasses at supplementary feeding stations (SFS), along with their level of antimicrobial resistance (AMR). E. coli was isolated from 90 (86.5%) samples but no ETEC was detected, likely because ETEC fimbriae confer the species specificity of the pathogen. Resistance to at least one antimicrobial agent was detected in 89.9% of E. coli isolates, being AMR levels extremely high (>70%) for tetracycline and streptomycin, and very high (>50%) for ampicillin and sulfamethoxazole-trimethoprim. Resistance to other critically important antimicrobials such as colistin and extended-spectrum cephalosporins was 2.2%, and 1.1%, respectively, and was encoded by the mcr-1 and blaSHV-12 genes. Multidrug resistance was displayed by 80% of the resistant E. coli and blaSHV-12 gene shared plasmid with other AMR genes. In general, resistance patterns in E. coli from vultures mirrored those found in pigs. C. difficile was detected in three samples (2.9%), two of them belonged to PCR-ribotype 078 and one to PCR-ribotype 126, both commonly found in pigs. All C. difficile isolates were characterized by a moderate to high level of resistance to fluoroquinolones and macrolides but susceptible to metronidazole or vancomycin, similar to what is usually found in C. difficile isolates from pigs. Thus, vultures may contribute somewhat to the environmental dissemination of some pig pathogens through their acquisition from pig carcasses and, more importantly, of AMR for antibiotics of critical importance for humans. However, the role of vultures would likely be much lesser than that of disposing pig carcasses at the SFS. The monitoring of AMR, and particularly of colistin resistant and ESLB-producing E. coli, should be considered in pig farms used as sources of carcasses for SFS.

[EN] Wild birds have repeatedly been found to be involved in the dissemination of enteric bacterial pathogens in the environment. The aim of this study was to determine the occurrence of Salmonella and Campylobacter as well as the antimicrobial resistance in wild Bonelli's eagles nestlings in Eastern Spain. In addition, we compared the efficiency of two sampling methods (fresh faecal samples from nest and cloacal swabs from nestlings) for detection of both bacteria. A total of 28 nests with 45 nestlings were analysed. In the nest, Salmonella occurrence was 61 +/- 9.2%, while Campylobacter occurrence was 11 +/- 5.8% (p < 0.05). In the nestlings, Salmonella occurrence was 36 +/- 7.1%, while Campylobacter occurrence was 11 +/- 4.7% (p < 0.05). Eight Salmonella serovars were identified, and the most frequently isolated were S. Enteritidis, S. Typhimurium, S. Houston, and S. Cerro. Only one Campylobacter species was identified (C. jejuni). Regarding antimicrobial resistance, the Salmonella strains isolated were found to be most frequently resistant to ampicillin and to tigecycline; however, the sole Campylobacter strain recovered was multidrug resistant. In conclusion, this study demonstrated that wild Bonelli's eagles nestlings are greater carriers of Salmonella than of Campylobacter. Both Salmonella and Campylobacter isolates exhibited antimicrobial resistance. In addition, faecal samples from nests were most reliable for Salmonella detection, while cloacal swab from nestlings were most reliable for Campylobacter detection., We wish to thank the Ministry of Infrastructures, Territory and Environment (Regional Government/Generalitat Valenciana), the research group "Improvement of Production System-related Food Safety and End Products" research group (Veterinary Faculty, University CEU-Cardenal Herrera) and GEMAS (Study Group on Wildlife Medicine and Conservation) for their technical support. Moreover, we want to thank University CEU-UCH (Consolidacion de Indicadores INDI 18/19 and IDOC 18/12) for the financial support. The English text version was revised by N. Macowan English Language Service.

Summary Phage-inducible chromosomal islands (PICIs) represent a novel and universal class of mobile genetic elements, which have broad impact on bacterial virulence. In spite of their relevance, how the Gram-negative PICIs hijack the phage machinery for their own specific packaging and how they block phage reproduction remains to be determined. Using genetic and structural analyses, we solve the mystery here by showing that the Gram-negative PICIs encode a protein that simultaneously performs these processes. This protein, which we have named Rpp (for redirecting phage packaging), interacts with the phage terminase small subunit, forming a heterocomplex. This complex is unable to recognize the phage DNA, blocking phage packaging, but specifically binds to the PICI genome, promoting PICI packaging. Our studies reveal the mechanism of action that allows PICI dissemination in nature, introducing a new paradigm in the understanding of the biology of pathogenicity islands and therefore of bacterial pathogen evolution., Graphical Abstract, Highlights • PICI Rpp protein promotes PICI transfer while blocking phage reproduction • Rpp forms a heterocomplex with helper phage TerS • Crystal structures of Rpp alone or complexed with TerS were determined • TerS complexed with Rpp switches specificity from the phage DNA to the PICI genome, Fillol-Salom et al. report that Gram-negative PICIs employ the Rpp protein to block helper phage reproduction. Rpp binds to helper phage TerS, and the Rpp-TerS heterocomplex enables PICIs to hijack the phage machinery for their own packaging. These findings reveal a mechanism used by Gram-negative PICIs to spread in nature.

Pseudomonas putida are ubiquitous inhabitants of soils and clinical isolates of this species have been seldom described. Clinical isolates show significant variability in their ability to cause damage to hosts because some of them are able to modulate the host’s immune response. In the current study, comparisons between the genomes of different clinical and environmental strains of P. putida were done to identify genetic clusters shared by clinical isolates that are not present in environmental isolates. We show that in clinical strains specific genes are mostly present on transposons, and that this set of genes exhibit high identity with genes found in pathogens and opportunistic pathogens. The set of genes prevalent in P. putida clinical isolates, and absent in environmental isolates, are related with survival under oxidative stress conditions, resistance against biocides, amino acid metabolism and toxin/antitoxin (TA) systems. This set of functions have influence in colonization and survival within human tissues, since they avoid host immune response or enhance stress resistance. An in depth bioinformatic analysis was also carried out to identify genetic clusters that are exclusive to each of the clinical isolates and that correlate with phenotypical differences between them, a secretion system type III-like was found in one of these clinical strains, a determinant of pathogenicity in Gram-negative bacteria., Work in the authors’ laboratories was supported by ERANET Pathogenomics programme through the ADHERS project (Ref: BIO2008-04419-E) and Fondos FEDER from the European Union through project BIO2010-17227 of the Ministry of Economy and Competitivity of Spain. Bio-Iliberis R&D provided support

La caracterización de bacterias ácido lácticas del género Lactobacillus aisladas y seleccionadas de líquido ruminal vacuno en el Municipio Colon del estado Zulia, se llevó a cabo mediante el muestreo de animales post-morten en el centro de matanza y servicio de distribución de carnes “FIBASA”. Dichas muestras fueron diluidas a diferentes concentraciones desde 10-1 – 10-7 y sembradas en placas, obteniendo con ello colonias bacterianas, las cuales fueron evaluadas in-vitro con pruebas de crecimiento, forma, color, tamaño, movilidad, presencia de esporas, producción de ácido láctico, fermentación de carbohidratos, resistencia a ácidos (2,5 – 6,5) y perfiles térmicos desde (4°C – 50°C). Con ello se logró caracterizar dos cepas (L1 y L2), cuyo análisis bioquímico y morfológico las ubica dentro del género Lactobacillus, estas cepas presentaron una alta actividad antagónica contra bacterias patógenas, los que las convierte en un potencial probiótico para la alimentación animal.

For nearly 3 decades, listeriologists and immunologists have used mainly three strains of the same serovar (1/2a) to analyze the virulence of the bacterial pathogen Listeria monocytogenes. The genomes of two of these strains, EGD-e and 10403S, were released in 2001 and 2008, respectively. Here we report the genome sequence of the third reference strain, EGD, and extensive genomic and phenotypic comparisons of the three strains. Strikingly, EGD-e is genetically highly distinct from EGD (29,016 single nucleotide polymorphisms [SNPs]) and 10403S (30,296 SNPs), and is more related to serovar 1/2c than 1/2a strains. We also found that while EGD and 10403S strains are genetically very close (317 SNPs), EGD has a point mutation in the transcriptional regulator PrfA (PrfA*), leading to constitutive expression of several major virulence genes. We generated an EGD-e PrfA*mutant and showed that EGD behaves like this strain in vitro, with slower growth in broth and higher invasiveness in human cells than those of EGD-e and 10403S. In contrast, bacterial counts in blood, liver, and spleen during infection in mice revealed that EGD and 10403S are less virulent than EGD-e, which is itself less virulent than EGD-e PrfA*. Thus, constitutive expression of PrfA-regulated virulence genes does not appear to provide a significant advantage to the EGD strain during infection in vivo, highlighting the fact that in vitro invasion assays are not sufficient for evaluating the pathogenic potential of L. monocytogenes strains. Together, our results pave the way for deciphering unexplained differences or discrepancies in experiments using different L. monocytogenes strains. © 2014 Bécavin et al., European Research Council (advanced grant 233348), the French Agence Nationale de la Recherche (grants BACNET 10-BINF-02-01, IBEID ANR-10-LABX-62-01, and ERA-NET ANR-2010-PATH), the Institut Pasteur, the Institut National de la Santé et de la Recherche Médicale, and the Institut National de la Recherche Agronomique. A.K