Center of Collecting, Welcome and Handling of Wild Animals (CRATAS), University of Trá s-os-Montes and Alto Douro, Vila Real, PortugalA total of 36 Escherichia coli and 31 enterococci isolates were recovered from 42 common buzzard faecal samples. The E. coli isolates showed high levels of resistance to streptomycin and tetracycline. The following resistance genes were detected: bla TEM (20 of 22 ampicillin-resistant isolates), tet(A) and/or tet(B) (16 of 27 tetracycline-resistant isolates), aadA1 (eight of 27 streptomycin-resistant isolates), cmlA (three of 15 chloramphenicol-resistant isolates), aac(3)-II with/without aac(3)-IV (all seven gentamicin-resistant isolates) and sul1 and/or sul2 and/or sul3 [all eight sulfamethoxazole/trimethoprim-resistant (SXT) isolates]. intI1 and intI2 genes were detected in four SXT-resistant isolates. The virulence-associated genes fimA (type 1 fimbriae), papC (P fimbriae) and aer (aerobactin) were detected in 61.1, 13.8 and 11.1 % of the isolates, respectively. The isolates belonged to phylogroups A (47.2 %), B1 (8.3 %), B2 (13.9 %) and D (30.5 %). For the enterococci isolates, Enterococcus faecium was the most prevalent species (48.4 %). High levels of tetracycline and erythromycin resistance were found among our isolates (87 and 81 %, respectively). Most of the tetracycline-resistant strains carried the tet(M) and/or tet(L) genes. The erm(B) gene was detected in 80 % of erythromycin-resistant isolates. The vat(D) and/or vat(E) genes were found in nine of the 17 quinupristin-dalfopristin-resistant isolates. The enterococcal isolates showing high-level resistance for kanamycin, gentamicin and streptomycin contained the aph(39)-IIIa, aac(69)-aph(20) and ant(6)-Ia genes, respectively. This report reveals that common buzzards seem to represent an important reservoir, or at least a source, of multiresistant E. coli and enterococci isolates, and consequently may represent a considerable hazard to human and animal health by transmission of these isolates to waterways and other environmental sources via their faecal deposits.

To study the prevalence of vancomycin-resistant enterococci (VRE) and extended-spectrum b-lactamase (ESBL)-containing Escherichia coli isolates, and the mechanisms of resistance implicated, 220 faecal samples from wild birds were collected between 2006 and 2010 in the Azores Archipelago. Samples were spread on SlanetzÁBartley agar plates supplemented with 4 mg/l vancomycin and on Levine agar plates supplemented with 2 mg/l cefotaxime for VRE and ESBL-containing E. coli isolation, respectively. vanA-containing enterococcal isolates (four Enterococcus faecium and two Enterococcus durans) and vanC-1 Enterococcus gallinarum isolates were detected in six and seven faecal samples, respectively. VRE isolates showed ampicillin (n 011), ciprofloxacin (n 09), tetracycline (n06), erythromycin (n 05), quinupristin/dalfopristin (n 03) and high-level kanamycin resistance (n 01). The tet(L) and/or tet(M) gene was found in all tetracycline-resistant isolates and the erm(B) gene in all erythromycin-resistant isolates. Three vanAcontaining E. faecium and two E. gallinarum presented specific sequences of the Tn5397 transposon. Four VRE isolates harboured the ace virulence gene. One faecal sample revealed one ESBL-containing E. coli isolate that belongs to the A phylogenetic group, showed a phenotype of resistance to b-lactams and tetracycline, and harboured the bla CTX-M-14 , bla SHV-12 and the tet(A) genes. To our knowledge, this is the first study to focus on defining the prevalence of VRE and/or ESBL-containing E. coli strains in wild birds from the Azores. The data recovered are essential to improve knowledge about the dissemination of resistant strains through wild ecosystems and their possible implications by transferring these resistances to other animals or to humans.

Significance and Impact of the Study: This study shows antimicrobial resistance in commensal bacteria from the free-range, Portuguese, Iberian wolf population. The results indicate that the Iberian wolf could contribute to the spread of resistant bacteria throughout the environment. Additionally, in case of infection, an increased risk of therapeutic failure due to the presence of multiresistant bacteria may represent a health problem for this endangered species. Future studies must be performed to analyse the possible contamination of these animals through the environment and/or the food chain. AbstractThe aim of this study was to report the antimicrobial resistance, the molecular mechanisms associated and the detection of virulence determinants within faecal Enterococcus spp. and Escherichia coli isolates of Iberian wolf. Enterococci (n = 227) and E. coli (n = 195) isolates were obtained from faecal samples of Iberian wolf (Canis lupus signatus). High rates of resistance were detected for tetracycline and erythromycin among the enterococci isolates, and most of resistant isolates harboured the tet(M) and/or tet(L) and erm(B) genes, respectively. The bla TEM, tet(A) and/or tet(B), and aadA or strA-strB genes were detected among most ampicillin-, tetracycline-or streptomycin-resistant E. coli isolates, respectively. E. coli isolates were ascribed to phylogroups A (n = 56), B1 (91), B2 (13) and D (35). The occurrence of resistant enterococci and E. coli isolates in the faecal flora of Iberian wolf, including the presence of resistant genes in integrons, and virulence determinants was showed in this study. Iberian wolf might act as reservoir of certain resistance genes that could be spread throughout the environment.

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a b s t r a c tThis paper discusses the overall energy consumption and greenhouse gas emissions when extracting pigments and producing hydrogen from Spirogyra sp. microalga biomass. The energy evaluation from the biomass leftovers was also included in this work. The influence of the functional unit and different allocation criteria on the biorefinery assessments is also shown. The study consists of laboratory tests showing Spirogyra sp. growth, harvesting, drying, pigment extraction and fermentation by Clostridium butyricum. Electrocoagulation and solar drying were tested and compared to conventional centrifugation and electrical dewatering in terms of their energy consumption for harvesting and dewatering, respectively. To discuss the biorefinery viability, the pigments and biohydrogen (bioH 2 ) retail costs are considered against operational costs according to electricity needs. The low yield of biochemical hydrogen and the high energy requirements for the pigment extraction were identified as main topics for further research. This research hopefully contributes to highlight the importance of energy and emission balances in order to decide on feasibility of the biorefinery.

In order to achieve stringent environmental and safety requirements, refineries are in search of "green" and cost-effective methods for crude oil desulphurization. Combined desulphurization technologies are being studied, including bioprocessing to upgrade fossil fuels. Using biodesulphurization (BDS), which is a biochemical process mediated by specific microorganisms, it is possible to desulphurize most of the hydrodesulphurization (HDS) recalcitrant sulphur compounds under mild operating conditions, making it a simple and eco-friendly process. In this study, two BDS process designs are compared, in terms of energy consumption, greenhouse gas emissions and operational costs by following a life cycle assessment (LCA) and life cycle cost (LCC) based methodology. The industrial HDS process is used as the reference technology for sulphur removal from fossil fuels. Different theoretical scenarios were considered and the best BDS results are scaled-up to evaluate a case study of providing ultra low sulphur diesel to an urban taxi fleet. This study exploits the potential of BDS as a cost-effective and ecofriendly alternative or complementary technology to the commonly HDS towards ultra low sulphur fuels.

a b s t r a c tAn alternative to the sugar/starch-based crops bioethanol is lignocellulosic biomass, but its utilization to biofuels is still not economically viable. In this context, an increasing interest has arising on the search for specific energy crops that do not require arable lands and are not water intensive, such as Jerusalem artichoke (JA). So, this work consisted on the cultivation of JA on those agricultural conditions and its further evaluation as a sustainable feedstock towards bioethanol. Two strategies of producing bioethanol were evaluated pointing out for the consolidated bioprocessing with the Zygosaccharomyces bailii Talf1 yeast as the best approach for further scale-up, based on energy data analysis and ethanol productivity. Different industrial scenarios were outlined and compared for overall CO 2 eq emissions and energy consumption per liter of ethanol (L EtOH ), using adequate criteria on a cradle-to-gate approach. With no land-use change, no biogenic and no co-products credits, the comparison of the overall energy consumption and CO 2 eq emissions (100% process) from JA ethanol (9 MJ/L EtOH ; 679 g CO 2 /L EtOH ) with sugarcane/sugar beet ethanol (42/29 MJ/L EtOH ; 731/735 g CO 2 /L EtOH ) and with gasoline refinery (15 MJ/L EtOH eq ; 1154 g CO 2 /L EtOH eq ), highlights the JA as an alternative feedstock to be a focus of ethanol research for gasoline blends.