Detection of vanA-containing Enterococcus species in faecal microbiota of gilthead seabream (Sparus aurata)

Vancomycin-resistant Enterococcus faecalis, E. faecium and E. durans isolates with the genotype vanA were detected in 7 of 118 faecal samples (5.9%) of natural gilthead seabream recovered off the coast of Portugal, and one vancomycinresistant isolate/sample was further characterized. The genes erm(B), tet(L), tet(M), aac(6')-aph(2"), aph(3')-IIIa and/ or ant(6)-Ia were identified in most of the 7 vancomycin-resistant enterococci. Sequence types ST273, ST313 and ST76 were detected in three E. faecium isolates and ST6 in two E. faecalis isolates. VanA-containing enterococci are suggested to be disseminated in fish in marine ecosystems close to areas of human activity

Detection and genetic characterisation of vanA-containing enterococcus strains in healthy lusitano horses

Lusitano horses were investigated in order to detect the presence of vancomycin-resistant enterococci. vanA isolates showed high level vancomycin (Minimum inhibitory concentration; MIC 128 mg/l) and teicoplanin resistance (MIC 64 mg/l), as well as resistance to ciprofloxacin, erythromycin and tetracycline. The tet(L) and erm(B) genes, associated with tetracycline and erythromycin resistance, respectively, were found in all vanA isolates.The intestinal tract of Lusitano horses can be a potential reservoir for vanA-containing enterococci. © 2009 EVJ Ltd

Proteomic study in an Escherichia coli strain from seagulls of the berlengas natural reserve of Portugal

The increasing bacterial resistance among common pathogens is threatening the effectiveness of several antibiotics. This represents a serious public health problem as such bacterial strains have already been detected in domestic, wild-life animals and humans. Using Escherichia coli as a model organism, we applied a proteomic approach to the topic of antimicrobial resistance. In order to identify and characterize the proteome of extendedspectrum -lactamase (ESBL) type TEM-52 producing-Escherichia coli strain of a faecal sample taken from Yellow-legged seagulls (Larus cachinnans) a bidimensional electrophoresis (2-DE) technique with an isoelectric focusing followed by a SDS-PAGE, was used. Eighty seven individualized protein spots were identified. All were suitable for peptide mass fingerprinting by a mass spectrometric technique (MALDI/TOF MS). Their identification was carried out by searching appropriate bioinformatic databases. All proteins were related to E. coli strains. Detection of proteins related to several E. coli strains linked with virulent and enterohaemorrhagic consequences in ESBL producing-E. coli isolates of seagull samples raises the question of how such similarities arise bearing in mind these remarkably different microbial ecosystems. © 2011, Proteomass Scientific Society. All rights reserved

Unveiling the physicochemical properties of a sulfated polysaccharide based on Ulvan with high biomedical potential

Introduction: Ulvan, extracted from the green algae Ulva lactuca displays physico-chemical and biological features of potential interest for biomedical applications1. This sulfated polysaccharide is mainly built on disaccharides repeating sequences composed of sulphated rhamnose and glucuronic acid, iduronic acid or xylose2. In this work, structural, chemical degradation and rheological studies were performed in order to complement in vitro and in vivo performance studies and to screen its potential biomedical interest

Kefiran cryogels as potential scaffolds for drug delivery and tissue engineering applications

A Kefiran-based scaffold was developed using a freeze gelation technique, and its potential use for both drug delivery system and tissue engineering applications were investigated. This scaffold showed, through micro-computed tomography and scanning electron microscopy, high porosity (82.3 ± 4.4%), thick pore walls (13.4 ± 0.7 μm), aerogel form with foam-like structure, and elastic behaviour (δ = 16 ± 0.7°). Moreover, the Kefiran scaffold degradation showed a delayed profile for over 28 days. The developed scaffold allowed a slow and sustained diclofenac release over two weeks, and the human Adipose-derived Stem Cells, cultured onto the Kefiran scaffold, were metabolically active after 72 h. Therefore, our research suggests that Kefiran cryogel could be a potential candidate for drug delivery of controlled bioactive molecules and tissue engineering scaffolding.Hajer Radhouani, Cristiana Gonçalves and F. Raquel Maia were supported by the Fundação para a ciência e a Tecnologia (FCT) from Portugal, with references CEECIND/00111/2017, SFRH/BPD/94277/2013 and SFRH/BPD/117492/2016, respectively. Diana Bicho was supported through the M-ERA-NET/0001/2014 project (FCT). Joaquim. M. Oliveira would like to thank the FCT for the fund provided under the program Investigador FCT 2015 (IF/01285/2015). This work was funded by the R&D Project KOAT – Kefiran exopolysaccharide: Promising biopolymer for use in regenerative medicine and tissue engineering, with reference PTDC/BTMMAT/29760/2017 (POCI-01-0145-FEDER-029760), financed by FCT and co-financed by European Regional Development Fund (FEDER) and Operational Programme for Competitiveness and Internationalisation (POCI)

Iberian Lynx (Lynx pardinus) from the captive breeding program as reservoir of antimicrobial resistant enterococci and Escherichia coli isolates

A total of 98 faecal samples from captive specimens of Iberian lynx were collected and analysed for enterococci (96 isolates) and Escherichia coli (90 isolates) recovery. These 186 isolates were tested for antimicrobial resistance, molecular mechanisms of resistance, and detection of virulence genes. Among the enterococci, Enterococcus hirae was the most prevalent species (35 isolates), followed by Enterococcus faecalis (30 isolates), Enterococcus faecium (27 isolates), and Enterococcus durans (4 isolates). High rates of resistance to tetracycline, erythromycin and high-level-kanamycin were detected among enterococcal isolates (41%, 26%, and 19%, respectively). The tet(M) and/or tet(L), erm(B), aac(6)-Ie-aph(2)-Ia, ant(6)-Ia, or aph(3)-IIIa genes were detected among resistant enterococci. Likewise, high rates of resistance were detected in E. coli isolates to tetracycline, streptomycin, sulfamethoxazole-trimethoprim (SXT), nalidixic acid, ampicillin, and ciprofloxacin (34%, 28%, 26%, 21%, 17%, and 16%, respectively). Furthermore, the blaTEM or blaSHV, tet(A) and/or tet(B), aadA or strA-strB, aac(3)-II and/or aac(3)-IV, and different combinations of sul genes were detected among most resistant isolates. Fifteen isolates contained class 1 and/or class 2 integrons and 3 different gene cassette arrays were identified (aadA1, dfrA1+aadA1, and estX+psp+aadA2). The E. coli isolates were ascribed to phylo-groups A (12%); B1 (40%); B2 (37%), and D (11%), being fimA the most prevalent virulence gene (n=84), followed by aer (n=13), cnf1 (n=13), papC (n=10) and papG-allele III (n=9). This study showed specimens of Iberian lynx acting as reservoirs of resistance genes, and in future (re)introductions they could spread resistant bacteria throughout the environmen

Wild boars as reservoirs of extended-spectrum beta-lactamases in Escherichia coli isolates of the A, B1 and B2 phylogenetic groups

ESBL-producing E. coli isolates have been isolated from eight of seventy seven faecal samples (10.4%) of wild boars in Portugal. The ESBL types identified by PCR and sequencing were blaCTX-M-1 (6 isolates) and blaCTX-M-1 + blaTEM1-b (2 isolates). Further resistance genes detected included tet(A) or tet(B) (in three tetracycline-resistant isolates), aadA (in three streptomycin- resistant isolates), cmlA (in one chloramphenicol-resistant isolate), sul1 and/or sul2 and/or sul3 (in all sulfonamide-resistant isolates). The intI1 gene encoding class 1 integrase was detected in all ESBL-producing E. coli isolates. One isolate also carried the intI2 gene, encoding class 2 integrase. The ESBL-producing E. coli isolates could be assigned to phylogenetic groups B1 (3 isolates), B2 (3 isolates) or A (2 isolates). Amino acid change in GyrA protein (Ser83Leu or Asp87Tyr) was detected in three nalidixic acid-resistant and ciprofloxacin-susceptible isolates. Two amino acid changes in GyrA (Ser83Leu + Asp87Asn) and one in ParC (Ser80Ile) were identified in two nalidixic acid- and ciprofloxacin-resistant isolates. As evidenced by this study wild boars could be a reservoir of antimicrobial resistance genes. © 2009 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

Antimicrobial resistance in faecal Escherichia coli isolates from farmed red deer and wild small mammals. Detection of a multiresistant E. coli producing extended-spectrum beta-lactamase

Eighty-nine Escherichia coli isolates recovered from faeces of red deer and small mammals, cohabiting the same area, were analyzed to determine the prevalence and mechanisms of antimicrobial resistance and molecular typing. Antimicrobial resistance was detected in 6.7% of isolates, with resistances to tetracycline and quinolones being the most common. An E. coli strain carrying blaCTX-M-1 as well as other antibiotic resistant genes included in an unusual class 1 integron (Intl1-dfrA16-blaPSE-1-aadA2-cmlA1-aadA1-qacH-IS440-sul3-orf1-mef(B)-IS26) was isolated from a deer. The blaCTX-M-1 gene was transferred by conjugation and transconjugants also acquired an IncN plasmid. This strain was typed as ST224, which seems to be well adapted to both clinical and environmental settings. The phylogenetic distribution of the 89 strains varied depending on the animal host. This work reveals low antimicrobial resistance levels among faecal E. coli from wild mammals, which reflects a lower selective pressure affecting these bacteria, compared to livestock. However, it is remarkable the detection of a multi-resistant ESBL-E. coli with an integron carrying clinically relevant antibiotic-resistance genes, which can contribute to the dissemination of resistance determinants among different ecosystems. © 2016 Elsevier Ltd