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

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

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

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

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

High level resistance to gentamicin and vancomycin in enterococci isolated from poultry in Argentina

In chicken husbandry, antimicrobial resistance among enterococci has been observed. Glycopeptides and gentamicin MinimumInhibitory Concentrations in fecal poultry enterococci were determined. Glycopeptide and high-level gentamicin resistance(HLGR) genes were investigated. E. faecalis (n = 105) and E. faecium (n = 70) were the predominant species. In E. faecalis, HLGR(19%) was observed. In high-level gentamicin resistant E. faecalis, presence of aac (6´) -Ie-aph (2´´)-Ia gene was confirmed. VanA E.faecium (7.1%) were found, with HLGR (20%) and high-level streptomycin resistance (40%). vanA genotype was detected inglycopeptide resistant E. faecium (100%). These results highlight the presence of horizontally transferable antimicrobialresistance determinants in E. faecalis and E. faecium recovered from poultry. Existence of enterococcal reservoirs in poultry shouldbe taken into account as a potential source of antimicrobial resistance genes, strengthening their potential spread to humansthrough the food chain of horizontal transferable resistance determinants.Fil: 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: Sparo, Mónica Delfina. 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: Ceci, Monica. Centro de Estudios Bioquímicos, Tandil, Prov. Bs.as.; ArgentinaFil: Baldaccini, Maria Beatriz. Universidad Nacional del Centro de la Provincia de Buenos Aires; ArgentinaFil: Garcia Allende Leonardo. Centro de Estudios Bioquímicos, Tandil, Prov. Bs.as.; 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

Clinical and microbiological features of bacteremia caused by Enterococcus faecalis

Introduction: Enterococcus faecalis is a frequent etiologic agent of invasive infections in hospitalized patients. The aim of this study was to analyze clinical and microbiological features of bacteremia caused by E. faecalis. Methodology: Between 2011 and 2013, significant bacteremia caused by E. faecalis in hospitalized patients was studied. Patient characteristics, comorbid conditions, and 14-day mortality were recorded. Virulence genes esp, gelE, and cylA; opsonophagocytosis resistance; resistance to bactericidal effect of normal serum; beta lactamase production; and susceptibility to ampicillin, vancomycin, teicoplanin, gentamicin, and streptomycin were investigated. Results: E. faecalis strains were recovered from 33 bacteremic patients. Polymicrobial bacteremia was diagnosed in 2 patients; 10 patients died. Virulence genes were found in strains from both deceased patients and survivors. Sources of bacteremia included urinary tract infections (36.4%), vascular catheters (15.1%), abscesses (9.1%), and unknown (48.5%). Underlying diseases included cancer (30.3%), diabetes (36.4%), cirrhosis (6.1%), renal (36.4%), and chronic obstructive pulmonary disease (2.0%). Co-morbidities included alcohol use (26.1%); glucocorticoid therapy (19.0%); prior antibiotic therapy (60.6%); and central venous (21.2%), arterial (12.1%), and urinary (63.6%) catheters. Also, 57.6% of patients came from the intensive care unit (ICU); 33.3% had mechanical ventilation. Significant mortality-associated conditions included polymicrobial bacteremia, oncological disease, APACHE II score ≥ 20, ICU stay, renal disease, central venous catheter, and mechanical ventilation. Conclusions: Outcome of patients was associated with their status and not with the presence of virulence genes in E. faecalis strains. A significant percentage of bacteremia had undetermined origin. An alternate origin may be the gastrointestinal tract, through translocation.Facultad de Ciencias Médica

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.