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Callens B.F.,Ghent University | Haesebrouck F.,Ghent University | Maes D.,Ghent University | Butaye P.,CODA CERVA VAR | And 2 more authors.
Microbial Drug Resistance | Year: 2013

Streptococcus suis (S. suis) has often been reported as an important swine pathogen and is considered as a new emerging zoonotic agent. Consequently, it is important to be informed on its susceptibility to antimicrobial agents. In the current study, the Minimum Inhibitory Concentration (MIC) population distribution of nine antimicrobial agents has been determined for nasal S. suis strains, isolated from healthy pigs at the end of the fattening period from 50 closed or semiclosed pig herds. The aim of the study was to report resistance based on both clinical breakpoints (clinical resistance percentage) and epidemiological cutoff values (non-wild-type percentage). Non-wild-type percentages were high for tetracycline (98%), lincomycin (92%), tilmicosin (72%), erythromycin (70%), tylosin (66%), and low for florfenicol (0%) and enrofloxacin (0.3%). Clinical resistance percentages were high for tetracycline (95%), erythromycin (66%), tylosin (66%), and low for florfenicol (0.3%) and enrofloxacin (0.3%). For tiamulin, for which no clinical breakpoint is available, 57% of the isolates did not belong to the wild-type population. Clinical resistance and non-wild-type percentages differed substantially for penicillin. Only 1% of the tested S. suis strains was considered as clinically resistant, whereas 47% of the strains showed acquired resistance when epidemiological cutoff values were used. In conclusion, MIC values for penicillin are gradually increasing, compared to previous reports, although pigs infected with strains showing higher MICs may still respond to treatment with penicillin. The high rate of acquired resistance against tiamulin has not been reported before. Results from this study clearly demonstrate that the use of different interpretive criteria contributes to the extent of differences in reported antimicrobial resistance results. The early detection of small changes in the MIC population distribution of isolates, while clinical failure may not yet be observed, provides the opportunity to implement appropriate risk management steps. © Copyright 2013, Mary Ann Liebert, Inc. 2013. Source

Bjorkeng E.K.,University of Tromso | Tessema G.T.,University of Tromso | Tessema G.T.,Nofima Materials AS | Tessema G.T.,Norwegian University of Life Sciences | And 7 more authors.
Microbiology | Year: 2010

The presence, distribution and expression of cassette chromosome recombinase (ccr) genes, which are homologous to the staphylococcal ccrAB genes and are designated ccrABEnt genes, were examined in enterococcal isolates (n=421) representing 13 different species. A total of 118 (28 %) isolates were positive for ccrABEnt genes by PCR, and a number of these were confirmed by Southern hybridization with a ccrAEnt probe (n576) and partial DNA sequencing of ccrAEnt and ccrBEnt genes (n=38). ccrABEnt genes were present in Enterococcus faecium (58/216, 27 %), Enterococcus durans (31/38, 82 %), Enterococcus hirae (27/52, 50 %), Enterococcus casseliflavus (1/4, 25 %) and Enterococcus gallinarum (1/2, 50 %). In the eight other species tested, including Enterococcus faecalis (n=94), ccrABEnt genes were not found. Thirty-eight sequenced ccrAB Ent genes from five different enterococcal species showed 94-100% nucleotide sequence identity and linkage PCRs showed heterogeneity in the ccrABEnt flanking chromosomal genes. Expression analysis of ccrABEnt genes from the E. faecium DO strain showed constitutive expression as a bicistronic mRNA. The ccrABEnt mRNA levels were lower during log phase than stationary phase in relation to total mRNA. Multilocus sequence typing was performed on 39 isolates. ccrABEnt genes were detected in both hospital-related (10/29, 34 %) and non-hospital (4/10, 40 %) strains of E. faecium. Various sequence types were represented by both ccrABEnt positive and negative isolates, suggesting acquisition or loss of ccrABEnt in E. faecium. In summary, ccrABEnt genes, potentially involved in genome plasticity, are expressed in E. faecium and are widely distributed in the E. faecium and E. casseliflavus species groups. © 2010 SGM. Source

Doublet B.,French National Institute for Agricultural Research | Doublet B.,University of Tours | Praud K.,French National Institute for Agricultural Research | Praud K.,University of Tours | And 7 more authors.
Journal of Antimicrobial Chemotherapy | Year: 2014

Objectives: To characterize the genetic determinants responsible for extended-spectrum cephalosporin (ESC) resistance of D-tartrate-positive Salmonella enterica subsp. enterica serovar Paratyphi B (serovar Paratyphi B dT+) strains that have emerged in poultry and humans in Belgium during 2008-10. Methods: The ESC resistance genes among non-redundant serovar Paratyphi B dT+ strains were determined using PCR and sequencing. ESC phenotypes were horizontally transferred by conjugation. Extended-spectrum β-lactamase (ESBL)-or AmpC-carrying plasmids were typed by PCR-based replicon typing, plasmid multilocus sequence typing and restriction fragment length polymorphism. The genetic relationship of ESC-resistant strains was assessed by XbaI PFGE and multilocus sequence typing. Results: Since 2008, the proportion of serovar Paratyphi B dT+strains frombroiler origin has increased significantly to reach 36.5% in 2010. Among 95 non-duplicate serovar Paratyphi B dT+ strains, 35% were resistant to ESCs. At the same time, a few ESC-resistant serovar Paratyphi B dT+ strains from humans were also detected in Belgium. Themost prevalent ESBL gene, blaCTX-M-1, and the AmpC cephalosporinase gene blaCMY-2 were identified on various conjugative IncI1 plasmids of different sequence types and with different additional non-β-lactam phenotypes. Interestingly, the blaCTX-M-2 gene was located on large multireplicon IncHI2/P plasmids. In addition, highly ESCresistant strains contained both the ESBL CTX-M-2 and the AmpC CMY-2 encoded by the IncHI2/P and IncI1 plasmids, respectively. All ESC-resistant serovar Paratyphi B dT+strains belonged to sequence type 28 and showed the common PFGE pattern X8, as well as the chromosomal class 2 integron cassette array dfrA1-sat2-aadA1 previously described in the European poultry-associated serovar Paratyphi B dT+ clonal population. Conclusions: This study showed that the clonal population of multidrug-resistant serovar Paratyphi B dT+, persisting in broilers in Belgium for the last decade, recently acquired various plasmid-borne ESC resistance determinants, constituting a major concern for public health. Further surveillance programmes and research are an absolute necessity to understand their epidemiology and to propose interventions to limit the spread of ESC-and multidrug-resistant Salmonella spp.© The Author 2013. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. Source

Ferreira H.L.,University of Sao Paulo | Rauw F.,CODA CERVA VAR | Pirlot J.F.,CODA CERVA VAR | Reynard F.,Merial SAS | And 3 more authors.
Avian Pathology | Year: 2014

Avian influenza (AI) vaccines should be used as part of a whole comprehensive AI control programme. Vectored vaccines based on Newcastle disease virus (NDV) are very promising, but are so far licensed in only a few countries. In the present study, the immunogenicity and protection against a highly pathogenic H5N1 influenza challenge were evaluated after vaccination with an enterotropic NDV vector expressing an H5 haemagglutinin (rNDV-H5) in 1-day-old specific pathogen free chickens inoculated once, twice or once followed by a heterologous boost with an inactivated H5N9 vaccine (iH5N9). The heterologous prime/boost rNDV-H5/iH5N9 combination afforded the best level of protection against the H5N1 challenge performed at 6 weeks of age. Two rNDV-H5 administrations conferred a good level of protection after challenge, although only a cellular H5-specific response could be detected. Interestingly, a single administration of rNDV-H5 gave the same level of protection as the double administration but without any detectable H5-specific immune response. In contrast to AI immunity, a high humoral, mucosal and cellular NDV-specific immunity could be detected up to 6 weeks post vaccination after using the three different vaccination schedules. NDV-specific mucosal and cellular immune responses were slightly higher after double rNDV-H5 vaccination when compared with single inoculation. Finally, the heterologous prime/boost rNDV-H5/iH5N9 combination induced a broader detectable immunity including systemic, mucosal and cellular AI and NDV-specific responses. © 2014 Houghton Trust Ltd. Source

Ferreira H.L.,University of Sao Paulo | Vangeluwe D.,Royal Belgian Institute Of Natural Sciences | Van Borm S.,CODA CERVA VAR | Poncin O.,Royal Belgian Institute Of Natural Sciences | And 5 more authors.
Avian Diseases | Year: 2015

Homosubtypic and heterosubtypic immunity in mallards (Anas platyrhynchos) play an important role in the avian influenza virus (AIV) diversity. The mechanisms of AIV replication among wild birds and the role of immunity in AIV diversity have thus not been completely clarified. During the monitoring of AI circulation among wild waterfowl in 2007-2008, two viruses (H3N8 and H1N1) were isolated from ducks caught in a funnel trap located in La Hulpe wetland in Belgium. H3N8 viruses were revealed to be more prevalent in the mallard population than was H1N1, which might suggest a better adaptation to this species. In order to investigate this hypothesis, we characterized both isolated viruses biologically by experimental inoculation. Virus excretion and humoral response induced by both isolated viruses were evaluated in mallards after a first infection followed by a homo-or heterosubtypic reinfection under controlled experimental conditions. The H1N1 virus had a delayed peak of excretion of 4 days compared to the H3N8, but the virus shedding was more limited, earlier, and shorter after each reinfection. Moreover, the H3N8 virus could spread to all ducks after homo-or heterosubtypic reinfections and during a longer period. Although the humoral response induced by both viruses after infection and reinfection could be detected efficiently by competitive ELISA, only a minimal H1 antibody response and almost no H3-specific antibodies could be detected by the HI test. Our results suggest that the H3N8 isolate replicates better in mallards under experimental controlled conditions. © 2015 American Association of Avian Pathologists. Source

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