Draft genome sequence of enterococcus faecalis strain CECT7121, a corn silage isolate with antibacterial activity against gram-positive pathogens

Enterococcus faecalis CECT7121 is a corn silage probiotic bacterium that shows antibacterial activity against Gram-positive pathogens from different origins. Its genome sequence is 2.9 Mb long with a GC content of 37.3%. Genome annotation identified three bacteriocin gene clusters in the genome.Fil: Delpech, Gastón. Universidad Nacional del Centro de la Provincia de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Hebert, Elvira Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Sparo, Mónica Delfina. Universidad Nacional del Centro de la Provincia de Buenos Aires; ArgentinaFil: Saavedra, Maria Lucila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentin

Mobile Genetic Elements in Vancomycin-Resistant <em>Enterococcus faecium</em> Population

Horizontal gene transfer constitutes a key driving force in bacterial evolution. The ability to acquire mobile genetic elements encoding antimicrobial resistance has contributed to the emergence of Enterococcus faecium as one of the main human nosocomial opportunistic pathogens. The deep analysis of the vancomycin-resistant E. faecium (VREfm) population’s mobilome, as the architecture and evolution of the core genome enables to observe VREfm plasticity and power of adaptation in animals, plants, environment and food. The persistence of VREfm is facilitated by the exchange of plasmids, phages and conjugative transposons that have allowed them to achieve a rapid adaptation to changes in environmental conditions. They can acquire resistance determinants from several species and transfer resistance genes to other potentially pathogenic bacteria such as methicillin-resistant Staphylococcus aureus strains

Impact on public health of the spread of high-level resistance to gentamicin and vancomycin in enterococci

Antibiotic resistance has turned into a global public health issue. Enterococci are intrinsically resistant to many antimicrobials groups. These bacteria colonize dairy and meat products and integrate the autochthonous microbiota of mammal’s gastrointestinal tract. Over the last decades, detection of vanA genotype in Enterococcus faecium from animals and from food of animal origin has been reported. Vancomycin-resistant E. faecium has become a prevalent nosocomial pathogen. Hospitalized patients are frequently treated with broad-spectrum antimicrobials and this leads to an increase in the presence of VanA or VanB vancomycin-resistant enterococci in patients’ gastrointestinal tract and the risk of invasive infections. In humans, E. faecium is the main reservoir of VanA and VanB phenotypes. Acquisition of high-level aminoglycoside resistance is a significant therapeutic problem for patients with severe infections because it negates the synergistic effect between aminoglycosides and a cell-wall-active agent. The aac(60)-Ie-aph (200)-Ia gene is widely spread in E. faecalis and has been detected in strains of human origin and in the food of animal origin. Enzyme AAC(60)-Ie-APH(200)-Ia confers resistance to available aminoglycosides, except to streptomycin. Due to the fast dissemination of this genetic determinant, the impact of its horizontal transferability among enterococcal species from different origin has been considered. The extensive use of antibiotics in food-producing animals contributes to an increase in drug-resistant animal bacteria that can be transmitted to humans. Innovation is needed for the development of new antibacterial drugs and for the design of combination therapies with conventional antibiotics. Nowadays, semi-purified bacteriocins and probiotics are becoming an attractive alternative to the antibiotic in animal production. Therefore, a better understanding of a complex and relevant issue for Public Health such as high-level vancomycin and gentamicin resistance in enterococci and their impact is needed. Hence, it is necessary to consider the spread of vanA E. faecium and high-level gentamicin resistant E. faecalis strains of different origin in the environment, and also highlight the potential horizontal transferability of these resistance determinants to other bacteria.Fil: Sparo, Mónica Delfina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Delpech, Gastón. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: García Allende, Natalia. Hopital Alemán; Argentin

Enrofloxacin-based therapeutic strategy for the prevention of endometritis in susceptible mares

Enrofloxacin (EFX) is often used empirically to prevent uterine infections in mares in order to improve efficiency on Commercial Embryo Transfer Farms. This study investigated the uterine distribution of EFX and its metabolite ciprofloxacin (CFX) in mares and assessed the minimal inhibitory concentrations (MIC) of EFX against various common pathogens as a basis for establishing a rational dosing schedule. Plasma and uterine pharmacokinetic (PK) studies were performed in two groups (n = 5) of healthy mares following intravenous (i.v.) administration of EFX at either 2.5 and at 5 mg/kg bodyweight. Plasma and endometrial tissue samples, taken before for up to 48 h after treatment were analysed by Reverse Phase HPLC. MIC values for wild strains of Gram-negative (Escherichia coli, Pseudomonas aeruginosa) and Gram-positive bacteria (β-haemolytic streptococci) ranged from 0.25-2 and 1.5-3.0 μg/mL respectively. In terms of tissue distribution, the sum of the endometrial concentrations of the parent drug (EFX) and its active metabolite (CFX) (in terms of AUC), exceeded those in plasma by 249% and 941% following administration of EFX at 2.5 and 5 mg/kg respectively. After i.v. treatment with EFX at 5 mg/kg, endometrial concentrations of EFX and CFX above the MIC value were detected for 36-48 and 22-43 h posttreatment for Gram-negative and -positive isolates respectively. Concentrations above MIC were maintained for much shorter periods at the lower (2.5 mg/kg) treatment dose. Based on these results, a conventional dose (5 mg/kg) of EFX given prebreeding followed by two further doses at 36-48 h postbreeding are proposed as a rational strategy for using of EFX as a preventative therapy against a variety of common bacterial strains associated with equine endometritis.Fil: González, C. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias. Departamento de Fisiopatología. Laboratorio de Farmacología; ArgentinaFil: Moreno, L.. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias. Departamento de Fisiopatología. Laboratorio de Farmacología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Fumuso, E..Fil: García, J..Fil: Rivulgo, M..Fil: Confalonieri, A..Fil: Sparo, M..Fil: Sánchez Bruni, S.

Bacterial antagonist mediated protein molecules

Bacterial antagonism mediated by ribosomally synthesised peptides has gained considerable attention in recent years because of its potential applications in the control of undesirable microbiota. These peptides, generally referred to as bacteriocins, are defined as a heterogeneous group of ribosomally synthesised, proteinaceous substances (with or without further modifications) extracellularly secreted by many Gram-positive and some Gram-negative bacteria. Their mode of activity is primarily bactericidal and directed against closely related strains and species. These peptides are nearly all cationic and very often amphyphilic, which is reflected in the fact that many of these peptides kill their target cells by accumulation in the membrane causing increasing permeability and loss of barrier functions . Bacteriocins have been explored primarily as natural food preservatives, but there is much interest in exploring the application of these therapeutic peptides as antimicrobial agents since many of them exhibit antimicrobial activity against various important human pathogens.The suitability of bacteriocins such as pharmaceuticals is explored through measures of cytotoxicity, effects on the natural microbiota, and in vivo efficacy in mouse models. Bacteriocins are promising therapeutic agents.Fil: Urbizu, Lucia Paola. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias. Departamento de Fisiopatología. Laboratorio de Farmacología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tandil. Centro de Investigacion Veterinaria de Tandil; ArgentinaFil: Sparo, Mónica Delfina. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias. Departamento de Fisiopatología. Laboratorio de Farmacología; ArgentinaFil: Sanchez Bruni, Sergio Fabian. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias. Departamento de Fisiopatología. Laboratorio de Farmacología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tandil. Centro de Investigacion Veterinaria de Tandil; Argentin

Immunomodulatory properties of cell wall extract from Enterococcus faecalis CECT7121

The aim of this study was to investigate the immunomodulatory properties of cell wall extract from Enterococcus faecalis CECT7121, measuring the induction of cytokines TNF-α, IL-6, IL-10 and IL-12 from human peripheral blood mononuclear cells (PBMCs). Cell wall extract was prepared from their growth in brain heart infusion broth (18. h, 35. °C). Subsequently, toxicity of the obtained cell wall extract was tested in Balb-C mice. PBMCs were isolated from buffy coats at the Blood Transfusion Service of Hospital Ramón Santamarina (Tandil, Argentina). PBMCs were purified using standard Ficoll-Paque gradient centrifugation. Aliquots of purified leukocytes were incubated at 37. °C for 24. h with heat-killed E. faecalis CECT7121 and cell wall extract. Concentrations of IL-6, TNFα, IL-10 and IL-12 (p70) were measured by solid phase sandwich ELISA. Changes in appearance and behavior of mice were evidenced only in the group with the maximal concentration of wall cell extract used (10,000. μg). Cell wall extract and heat-killed E. faecalis CECT7121 induced the production of significantly higher amounts of Il-12, IL-6, TNF-α and IL-10 cytokines compared to the nonstimulated PBMCs. These findings provide helpful information on immunomodulation activity by cell wall extract in sight of the application of this compound in controlling certain infectious diseases.Facultad de Ciencias Médica

Antimicrobial Resistance of Uropathogenic Escherichia coli from Elderly Patients at a General Hospital, Argentina

Background:Urinary Tract Infection (UTI) is a common cause of morbidity and mortality in older adults.Objective:To investigate antimicrobial resistance of uropathogenic Escherichia coli from elderly patients in a General Hospital, Argentina.Method:During the period July 2011-July 2015, patients over 70 years old with urinary tract infections, without urinary catheters and with noantimicrobial therapy the previous week before sampling, were included. Phenotypic characterization was carried out. In vitroqualitative and quantitative antimicrobial resistances were investigated. Antimicrobials assayed: ampicillin, amoxicillin-clavulanate,cefazolin, cefuroxime, cefoxitin, cefotaxime, ceftazidime, cefepime, imipenem, ertapenem, gentamicin, nalidixic acid, ciprofloxacin,trimethoprim-sulfamethoxazole (TMS) and nitrofurantoin. Patients? medical records were produced, and risk factors were analyzedby multivariate analysis.Results:768 bacterial isolates were identified as E. coli. Resistances to ampicillin (80.5%), nalidixic acid (61.7%), ciprofloxacin (42.8%),TMS (37.6%), amoxicillin-clavulanate (28.6%), cefazolin (21.6%), cefuroxime (20.7%), gentamicin (13.8%), cefotaxime (9.7%),ceftazidime (9.7%), cefepime (8.4%), cefoxitin (3.1%) and nitrofurantoin (2.3%) were observed. Resistance to carbapenems was notexpressed. Production of extended spectrum β-lactamases was detected (7.6%) in community acquired (96%) and healthcareassociated (4%) isolates. The independent risk factors for urinary infections produced by multi-resistant E. coli were: diabetesmellitus, recurrent infections, hospitalization during the last year and exposure to β-lactams in the last 3 months. Conclusion:A high prevalence of resistance to β-lactams and to other antimicrobials was observed. Detection of antimicrobial multi-resistantisolates highlights the need of antimicrobial resistance surveillance in elderly patients with urinary tract infections.Fil: Delpech, Gastón. Universidad Nacional del Centro de la Provincia de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Garcia Allende, Natalia. Hospital Alemán; ArgentinaFil: Lissarrague, Sabina. Hospital Ramon Santamarina de Tandil; ArgentinaFil: Sparo, Mónica Delfina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentin

Immunomodulatory properties of cell wall extract from Enterococcus faecalis CECT7121

The aim of this study was to investigate the immunomodulatory properties of cell wall extract from Enterococcus faecalis CECT7121, measuring the induction of cytokines TNF-α, IL-6, IL-10 and IL-12 from human peripheral blood mononuclear cells (PBMCs). Cell wall extract was prepared from their growth in brain heart infusion broth (18. h, 35. °C). Subsequently, toxicity of the obtained cell wall extract was tested in Balb-C mice. PBMCs were isolated from buffy coats at the Blood Transfusion Service of Hospital Ramón Santamarina (Tandil, Argentina). PBMCs were purified using standard Ficoll-Paque gradient centrifugation. Aliquots of purified leukocytes were incubated at 37. °C for 24. h with heat-killed E. faecalis CECT7121 and cell wall extract. Concentrations of IL-6, TNFα, IL-10 and IL-12 (p70) were measured by solid phase sandwich ELISA. Changes in appearance and behavior of mice were evidenced only in the group with the maximal concentration of wall cell extract used (10,000. μg). Cell wall extract and heat-killed E. faecalis CECT7121 induced the production of significantly higher amounts of Il-12, IL-6, TNF-α and IL-10 cytokines compared to the nonstimulated PBMCs. These findings provide helpful information on immunomodulation activity by cell wall extract in sight of the application of this compound in controlling certain infectious diseases.Facultad de Ciencias Médica

Efficacy of the probiotic strain Enterococcus faecalis CECT7121 in diarrhoea prevention in newborn foals

The aim of this work was to assess the effectiveness of the probiotic strain Enterococcus faecalis CECT7121 for the prevention of neonatal diarrhea in one to three day-old foals. Fecal samples from twenty foals divided randomly in experimental and control groups were studied. The experimental group (n=10) received E. faecalis CECT7121 (1 x 1010 CFU/ml) orally in the first day of life and for a 6 days period. The control group (n=10) received distilled water for the same period. In 4/10 of the foals from the control group diarrhea was observed, conversely diarrhea cases were not observed in any of the foals belonging to the experimental group. The differences between groups were statistically significant (p&amp;lt;0.001). E. faecalis CECT7121 was isolated in faeces from the second day of treatment until six days after the end of the probiotic administration. Therefore, E. faecalis CECT 7121 may be considered as a biological tool for the prevention of neonatal diarrhea in foals. Any adverse effects, at least under the conditions of this assay, but more efficacy studies should be performed to evaluate this organism in the prevention or treatment of diseases in foals.El objetivo de este trabajo fue evaluar la eficacia del probiótico Enterococcus faecalis CECT7121 en la prevención de la diarrea neonatal en potrillos de uno a tres días de edad. Se estudiaron muestras de materia fecal de veinte potrillos divididas al azar en grupos experimental y control. Los animales del grupo experimental (n=10) recibieron E. faecalis CECT7121 (1 x 1010 UFC/ml) por vía oral en el primer día de vida y por un período de 6 días. El grupo control (n=10) recibió agua destilada por el mismo período. Se observó que el 40% de los potrillos del grupo control presentaron diarrea, mientras que no se observaron casos de diarrea en ninguno de los potrillos pertenecientes al grupo experimental. Las diferencias entre los grupos fueron estadísticamente significativas (p&amp;lt;0,001). E. faecalis CECT7121 se aisló en heces desde el segundo día de tratamiento hasta seis días después del final de la administración del probiótico. Por lo tanto este hallazgo indicaría que E. faecalis CECT7121 podría ser considerado como una herramienta biológica para la prevención de la diarrea neonatal en potrillos sin ningún efecto adverso, al menos bajo las condiciones del presente ensayo, aunque más estudios de eficacia se deberían realizar para evaluar este organismo en la prevención o el tratamiento de enfermedades de los potrillos en los haras

Efficacy of the probiotic strain Enterococcus faecalis CECT7121 in diarrhoea prevention in newborn foals

The aim of this work was to assess the effectiveness of the probiotic strain Enterococcus faecalis CECT7121 for the prevention of neonatal diarrhea in one to three day-old foals. Fecal samples from twenty foals divided randomly in experimental and control groups were studied. The experimental group (n=10) received E. faecalis CECT7121 (1 x 1010 CFU/ml) orally in the first day of life and for a 6 days period. The control group (n=10) received distilled water for the same period. In 4/10 of the foals from the control group diarrhea was observed, conversely diarrhea cases were not observed in any of the foals belonging to the experimental group. The differences between groups were statistically significant (p&lt;0.001). E. faecalis CECT7121 was isolated in faeces from the second day of treatment until six days after the end of the probiotic administration. Therefore, E. faecalis CECT 7121 may be considered as a biological tool for the prevention of neonatal diarrhea in foals. Any adverse effects, at least under the conditions of this assay, but more efficacy studies should be performed to evaluate this organism in the prevention or treatment of diseases in foals.