Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

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Molecular relatedness of ESBL/AmpC-producing Escherichia coli from humans, animals, food and the enviroment : a pooled analysis
Dorado-Garcia, Alejandro ; Smid, J.H. ; Pelt, Wilfrid Van; Bonten, M.J.M. ; Fluit, A.C. ; Bunt, Gerrita van den; Wagenaar, J.A. ; Hordijk, J. ; Dierikx, C.M. ; Veldman, K.T. ; Koeijer, A.A. de; Dohmen, W. ; Schmitt, H. ; Liakopoulos, A. ; Pacholewicz, Ewa ; Lam, T.J.G.M. ; Velthuis, Annet ; Heuvelink, A. ; Gonggrijp, Maaike ; Duijkeren, E. van; Hoek, A.H.A.M. van; Roda Husman, A.N. de; Blaak, H. ; Havelaar, A.H. ; Mevius, D.J. ; Heederik, D.J.J. - \ 2018
Journal of Antimicrobial Chemotherapy 73 (2018)2. - ISSN 0305-7453 - p. 339 - 347.
Background: In recent years, ESBL/AmpC-producing Escherichia coli ESBL/AmpC-EC) have been isolated with increasing frequency from animals, food, environmental sources and humans. With incomplete and scattered evidence, the contribution to the human carriage burden from these reservoirs remains unclear.
Objectives: To quantify molecular similarities between different reservoirs as a first step towards risk attribution.
Methods: Pooled data on ESBL/AmpC-EC isolates were recovered from 35 studies in the Netherlands comprising.27 000 samples, mostly obtained between 2005 and 2015. Frequency distributions of ESBL/AmpC genes from 5808 isolates and replicons of ESBL/AmpC-carrying plasmids from 812 isolates were compared across 22 reservoirs through proportional similarity indices (PSIs) and principal component analyses (PCAs).
Results: Predominant ESBL/AmpC genes were identified in each reservoir. PCAs and PSIs revealed close human–animal ESBL/AmpC gene similarity between human farming communities and their animals (broilers and pigs) (PSIs from 0.8 to 0.9). Isolates from people in the general population had higher similarities to those from human clinical settings, surface and sewage water and wild birds (0.7–0.8), while similarities to livestock or food reservoirs were lower (0.3–0.6). Based on rarefaction curves, people in the general population had more diversity in ESBL/AmpC genes and plasmid replicon types than those in other reservoirs.
Conclusions: Our ‘One Health’ approach provides an integrated evaluation of the molecular relatedness of ESBL/AmpC-EC from numerous sources. The analysis showed distinguishable ESBL/AmpC-EC transmission cycles in different hosts and failed to demonstrate a close epidemiological linkage of ESBL/AmpC genes and plasmid replicon types between livestock farms and people in the general population.
Samenvatting ESBL-Attributieanalyse (ESBLAT) : Op zoek naar de bronnen van antibioticaresistentie bij de mens
Mevius, Dik ; Heederik, Dick ; Duijkeren, Engeline ; Veldman, Kees ; Essen, Alieda van; Kant, Arie ; Liakopoulos, Apostolos ; Geurts, Yvon ; Pelt, Wilfrid van; Mughini Gras, Lapo ; Schmitt, Heike ; Dierikx, Cindy ; Hoek, Angela van; Evers, Eric ; Roda Husman, Annemaria de; Blaak, Hetty ; Dissel, Jaap van; Smid, Joost ; Dohmen, Wietske ; Dorado-Garcia, Alejandro ; Havelaar, Arie ; Hordijk, Joost ; Wagenaar, Jaap ; Fluit, Ad ; Bunt, Gerrita van den; Bonten, Marc ; Velthuis, Annet ; Heuvelink, Annet ; Buter, Rianne ; Gonggrijp, Maaike ; Santman-Berends, Inge ; Lam, Theo ; Urlings, Bert ; Heres, Lourens ; Bouwknecht, Martijn ; Groot, Jacques de - \ 2018
Netherlands : De Stichting TKI Agri&Food (TKI) - 11 p.
Rapport ESBL-Attributieanalyse (ESBLAT) : Op zoek naar de bronnen van antibioticaresistentie bij de mens
Mevius, Dik ; Heederik, Dick ; Duijkeren, Engeline ; Veldman, Kees ; Essen, Alieda van; Kant, Arie ; Liakopoulos, Apostolos ; Geurts, Yvon ; Pelt, Wilfrid van; Mughini Gras, Lapo ; Schmitt, Heike ; Dierikx, Cindy ; Hoek, Angela van; Evers, Eric ; Roda Husman, Annemaria de; Blaak, Hetty ; Dissel, Jaap van; Smid, Joost ; Dohmen, Wietske ; Dorado-Garcia, Alejandro ; Havelaar, Arie ; Hordijk, Joost ; Wagenaar, Jaap ; Fluit, Ad ; Bunt, Gerrita van den; Bonten, Marc ; Velthuis, Annet ; Heuvelink, Annet ; Buter, Rianne ; Gonggrijp, Maaike ; Santman-Berends, Inge ; Lam, Theo ; Urlings, Bert ; Heres, Lourens ; Bouwknecht, Martijn ; Groot, Jacques de - \ 2018
Netherlands : De Stichting TKI Agri&Food (TKI) - 73
Methicillin-resistant Staphylococcus pseudintermedius among dogs in the description of novel SCCmec variants
Duim, Birgitta ; Verstappen, Koen M.H.W. ; Kalupahana, Ruwani S. ; Ranathunga, Lakmali ; Fluit, Ad C. ; Wagenaar, Jaap A. - \ 2018
Veterinary Microbiology 213 (2018). - ISSN 0378-1135 - p. 136 - 141.
S. pseudintermedius - SCCmec variants - Sri Lanka - ΨSCCmec
The presence and genetic characteristics of methicillin-resistant Staphylococcus pseudintermedius (MRSP) in Sri Lanka was investigated to add additional insight into global spread, emergence and evolution of MRSP. A total of 234 samples from dogs visiting veterinary clinics were cultured for staphylococci and the genomes of the MRSP isolates were sequenced, to identify resistance genes, the multilocus sequence types (MLST) and spa types. From a questionnaire the history of antimicrobial treatment and patient information was obtained. S. pseudintermedius was isolated from 116/229 samples, eight of these were MRSP. Six MRSP CC45 isolates contained a pseudo-SCC element ΨSCCmec57395. Two isolates belonging to ST429 (CC761) and ST121 (CC121) contained novel variants of the SCCmec Type V(T) element. The elements were designated SCCmecV(T)SL/066, that carried additional transposon-related genes, and SCCmecV(T)SL/154 that carried a type III restriction-modification system, a type 7 ccr gene complex, and a cadA coding sequence. Thirty-seven percent of the dogs received antimicrobial treatment at the time of sampling of which four dogs were MRSP-positive. The proportion of MRSP among S. pseudintermedius is low compared to other countries, despite the fact that in Sri Lanka antimicrobials for treatment of dogs are available over the counter. Important is the finding of novel type V(T) SCCmec elements, which further underlines the high recombination frequency of SCC elements. The ΨSCCmec57395 was found in isolates of CC45, which is the only sequence type of MRSP known to contain this pseudo-cassette.
Staphylococcus aureus nasal colonization differs among pig lineages and is associated with the presence of other staphylococcal species
Verstappen, Koen M. ; Willems, Eveline ; Fluit, Ad C. ; Duim, Birgitta ; Martens, Marc ; Wagenaar, Jaap A. - \ 2017
Frontiers in Veterinary Science 4 (2017)JUN. - ISSN 2297-1769
Colonization - Methicillin-resistant S. aureus - Pigs - Staphylococci - Staphylococcus aureus
Staphylococcus aureus is a common colonizer in pigs, with methicillin-resistant S. aureus (MRSA) in particular being a potential health risk to humans. To reduce the exposure to humans, the colonization in pigs should be reduced. The aim of this study was to quantitatively compare the susceptibility of pig lineages for S. aureus colonization, and if the absence of S. aureus could be associated with the presence or absence of other staphylococcal species. Nasal samples (n = 129) were obtained from seven different pig lineages in the Netherlands, France, and Germany. S. aureus and other staphylococci were enumerated from these samples by real-time (RT)-PCR and culture. Associations were explored between the presence of S. aureus and other staphylococci. S. aureus was detected by RT-PCR on all farms and in samples from pigs of all lineages. Twenty-five percent of the pigs from lineage F (from two farms) were colonized with S. aureus, while in all other lineages it was more than 50% (p < 0.01). Moreover, in S. aureus-positive samples from pigs of lineage F smaller amounts of S. aureus were found than in other lineages. Staphylococcus sciuri, Staphylococcus cohnii, and Staphylococcus saprophyticus were usually not found in combination with S. aureus in these samples. In conclusion: (i) pigs from different genetic lineages have different susceptibilities for colonization with S. aureus. These pigs might contain a genetic factor influencing nasal colonization. (ii) Colonization of S. aureus is also associated with the absence of S. sciuri, S. cohnii, or S. saprophyticus. (iii) The farm environment seems to influence the presence of S. aureus in pigs.
Development of a real-time PCR for detection of Staphylococcus pseudintermedius using a novel automated comparison of whole-genome sequences
Verstappen, Koen M. ; Huijbregts, Loes ; Spaninks, Mirlin ; Wagenaar, Jaap A. ; Fluit, Ad C. ; Duim, Birgitta - \ 2017
PLoS One 12 (2017)8. - ISSN 1932-6203 - 10 p.

Staphylococcus pseudintermedius is an opportunistic pathogen in dogs and cats and occasionally causes infections in humans. S. pseudintermedius is often resistant to multiple classes of antimicrobials. It requires a reliable detection so that it is not misidentified as S. aureus. Phenotypic and currently-used molecular-based diagnostic assays lack specificity or are labour-intensive using multiplex PCR or nucleic acid sequencing. The aim of this study was to identify a specific target for real-time PCR by comparing whole genome sequences of S. pseudintermedius and non-pseudintermedius.Genome sequences were downloaded from public repositories and supplemented by isolates that were sequenced in this study. A Perl-script was written that analysed 300-nt fragments from a reference genome sequence of S. pseudintermedius and checked if this sequence was present in other S. pseudintermedius genomes (n = 74) and non-pseudintermedius genomes (n = 138). Six sequences specific for S. pseudintermedius were identified (sequence length between 300–500 nt). One sequence, which was located in the spsJ gene, was used to develop primers and a probe. The real-time PCR showed 100% specificity when testing for S. pseudintermedius isolates (n = 54), and eight other staphylococcal species (n = 43). In conclusion, a novel approach by comparing whole genome sequences identified a sequence that is specific for S. pseudintermedius and provided a real-time PCR target for rapid and reliable detection of S. pseudintermedius.

ESBL/AmpC-producing Enterobacteriaceae in households with children of preschool age: prevalence, risk factors and co-carriage
Bunt, G. van den; Liakopoulos, A. ; Mevius, D.J. ; Geurts, Y. ; Fluit, A.C. ; Bonten, M.J.M. ; Mughini-Gras, Lapo ; Pelt, W. van - \ 2016
Journal of Antimicrobial Chemotherapy 72 (2016)2. - ISSN 0305-7453 - p. 589 - 595.
Objectives ESBL/AmpC-producing Enterobacteriaceae are an emerging public health concern. As households with preschool children may substantially contribute to the community burden of antimicrobial resistance, we determined the prevalence, risk factors and co-carriage of ESBL/AmpC-producing bacteria in preschool children and their parents. Methods From April 2013 to January 2015, each month 2000 preschool children were randomly selected from Dutch population registries. The parents were invited to complete an epidemiological questionnaire and to obtain and send a faecal sample from the selected child and from one parent. Samples were tested for ESBL/AmpC-producing bacteria. Logistic regression was used to identify risk factors for ESBL/AmpC carriage in children and parents, and findings were internally validated by bootstrapping. Results In total, 1016 families were included and ESBL/AmpC prevalence was 4.0% (95% CI 3.2%–5.0%); 3.5% (95% CI 2.5%–4.8%) in children and 4.5% (95% CI 3.4%–6.0%) in parents. Attending a daycare centre (DCC) was the only significant risk factor for children (OR 2.1, 95% CI 1.0–4.3). For parents, the only significant risk factor was having one or more children attending DCCs (OR 2.2, 95% CI 1.2–4.8). For parents of ESBL/AmpC-positive children the OR for ESBL/AmpC carriage was 19.7 (95% CI 9.2–42.4). Co-carriage of specific ESBL/AmpC genotypes in child and parent occurred more often than expected by chance (14.6% versus 1.1%, P < 0.001). Conclusions In this study, intestinal carriage with ESBL/AmpCs was detected in ∼4% of households with preschool children. DCC attendance was a risk factor in both children and parents and co-carriage of specific genotypes frequently occurred in child–parent pairs. These findings suggest household transmission or/and family-specific exposure to common sources of ESBL/AmpC-producing bacteria.
The effectiveness of bacteriophages against methicillin-resistant Staphylococcus aureus ST398 nasal colonization in pigs
Verstappen, Koen M. ; Tulinski, Pawel ; Duim, Birgitta ; Fluit, Ad C. ; Carney, Jennifer ; Nes, Arie Van; Wagenaar, Jaap A. - \ 2016
PLoS One 11 (2016)8. - ISSN 1932-6203

Methicillin-resistant Staphylococcus aureus (MRSA) is an important colonizer in animals and an opportunistic pathogen in humans. In humans, MRSA can cause infections that might be difficult to treat because of antimicrobial resistance. The use of bacteriophages has been suggested as a potential approach for the control of MRSA colonization to minimize the - often occupational - exposure of humans. The aim of this study was to assess the efficacy of bacteriophage treatment on porcine nasal colonization with MRSA in vitro, in vivo, and ex vivo. The effectiveness of a bacteriophage combination of phage K∗710 and P68 was assessed in vitro by incubating them with MRSA V0608892/1 (ST398) measuring the OD600 hourly. To study the in vivo effect, bacteriophages were administered in a gel developed for human application, which contain 109 plaque-forming units (pfu)/mL (K and P68 in a 19.25:1 ratio) for 5 days to piglets (N = 8) that were experimentally colonized with the MRSA strain. Eight piglets experimentally colonized were used as a negative control. The MRSA strain was also used to colonize porcine nasal mucosa explants and bacteriophages were applied to assess the ex vivo efficacy of treatment. Bacteriophages were effective in vitro. In vivo, sixteen piglets were colonized with MRSA but the number of CFU recovered after the application of the bacteriophages in 8 piglets was not reduced compared to the control animals (approx. 105 CFU/swab). In the ex vivo model, 108 CFU were used to establish colonization with MRSA; a reduction of colonization was not observed after application of bacteriophages. However, application of mupirocin both in vivo and ex vivo resulted in a near eradication of MRSA. In conclusion: i) The MRSA strain was killed in the presence of the bacteriophages phage K∗710 and P68 in vitro. ii) Bacteriophages did not reduce porcine nasal colonization in vivo or ex vivo. Physiological in vivo and ex vivo conditions may explain these observations. Efficacy in the ex vivo model matched that of the in vivo system.

Staphylococcus aureus ST398 gene expression profiling during ex vivo colonization of porcine nasal epithelium
Tulinski, P. ; Duim, B. ; Wittink, F.R. ; Jonker, M.J. ; Breit, T.M. ; Putten, J.P. van; Wagenaar, J.A. ; Fluit, A.C. - \ 2014
BMC Genomics 15 (2014). - ISSN 1471-2164
clumping factor-b - methicillin-resistant - carriage - model - adherence - humans - proteinases - determinant - infections - cells
Background: Staphylococcus aureus is a common human and animal opportunistic pathogen. In humans nasal carriage of S. aureus is a risk factor for various infections. Methicillin-resistant S. aureus ST398 is highly prevalent in pigs in Europe and North America. The mechanism of successful pig colonization by MRSA ST398 is poorly understood. Previously, we developed a nasal colonization model of porcine nasal mucosa explants to identify molecular traits involved in nasal MRSA colonization of pigs. Results: We report the analysis of changes in the transcription of MRSA ST398 strain S0462 during colonization on the explant epithelium. Major regulated genes were encoding metabolic processes and regulation of these genes may represent metabolic adaptation to nasal mucosa explants. Colonization was not accompanied by significant changes in transcripts of the main virulence associated genes or known human colonization factors. Here, we documented regulation of two genes which have potential influence on S. aureus colonization; cysteine extracellular proteinase (scpA) and von Willebrand factor-binding protein (vWbp, encoded on SaPIbov5). Colonization with isogenic-deletion strains (Delta vwbp and Delta scpA) did not alter the ex vivo nasal S. aureus colonization compared to wild type. Conclusions: Our results suggest that nasal colonization with MRSA ST398 is a complex event that is accompanied with changes in bacterial gene expression regulation and metabolic adaptation.
Methicillin resistant coagulase-negative staphylococci on pig farms and as a reservoir of heterogeneous staphylococcal casette chromosome mec elements
Tulinski, P. ; Fluit, A.C. ; Wagenaar, J.A. ; Mevius, D.J. ; Vijver, L.P.L. van de; Duim, B. - \ 2012
Applied and Environmental Microbiology 78 (2012)2. - ISSN 0099-2240 - p. 299 - 304.
aureus - sccmec - strains - susceptibility - identification - diversity - evolution - region - sentry - pcr
Methicillin-resistant Staphylococcus aureus (MRSA) likely originated by acquisition of the staphylococcal cassette chromosome mec (SCCmec) from coagulase-negative staphylococci (CNS). However, it is unknown whether the same SCCmec types are present in MRSA and CNS that reside in the same niche. Here we describe a study to determine the presence of a potential mecA reservoir among CNS recovered from 10 pig farms. The 44 strains belonged to 10 different Staphylococcus species. All S. aureus strains belonged to sequence type 398 (ST398), with SCCmec types V and IVa. Type IVc, as well as types III and VI, novel subtypes of type IV, and not-typeable types, were found in CNS. S. aureus, S. epidermidis, and S. haemolyticus shared SCCmec type V. The presence of SCCmec type IVc in several staphylococcal species isolated from one pig farm is noteworthy, suggesting exchange of this SCCmec type in CNS, but the general distribution of this SCCmec type still has to be established. In conclusion, this study shows that SCCmec types among staphylococcal species on pig farms are heterogeneous. On two farms, more than one recovered staphylococcal species harbored the same SCCmec type. We conclude that staphylococci on pig farms act as a reservoir of heterogeneous SCCmec elements. These staphylococci may act as a source for transfer of SCCmec to S. aureus.
Dutch patients, retail chicken meat and poultry share the same ESBL genes, plasmids and strains
Leverstein-van Hall, M.A. ; Dierikx, C.M. ; Cohen Stuart, J. ; Voets, G.M. ; Munckhof, M.P. Van den; Essen-Zandbergen, A. Van; Platteel, T. ; Fluit, A.C. ; Sande-Bruinsma, N. Van de; Scharinga, J. ; Bonten, M.J.M. ; Mevius, D.J. - \ 2011
Clinical Microbiology and Infection 17 (2011)6. - ISSN 1198-743X - p. 873 - 880.
spectrum beta-lactamases - resistant escherichia-coli - extended-spectrum - salmonella-enterica - humans - infections - prevalence - diversity - emergence - trends
Intestinal carriage of extended-spectrum beta-lactamase (ESBL) -producing bacteria in food-producing animals and contamination of retail meat may contribute to increased incidences of infections with ESBL-producing bacteria in humans. Therefore, distribution of ESBL genes, plasmids and strain genotypes in Escherichia coli obtained from poultry and retail chicken meat in the Netherlands was determined and defined as ‘poultry-associated’ (PA). Subsequently, the proportion of E. coli isolates with PA ESBL genes, plasmids and strains was quantified in a representative sample of clinical isolates. The E. coli were derived from 98 retail chicken meat samples, a prevalence survey among poultry, and 516 human clinical samples from 31 laboratories collected during a 3-month period in 2009. Isolates were analysed using an ESBL-specific microarray, sequencing of ESBL genes, PCR-based replicon typing of plasmids, plasmid multi-locus sequence typing (pMLST) and strain genotyping (MLST). Six ESBL genes were defined as PA (blaCTX-M-1, blaCTX-M-2, blaSHV-2, blaSHV-12, blaTEM-20, blaTEM-52): 35% of the human isolates contained PA ESBL genes and 19% contained PA ESBL genes located on IncI1 plasmids that were genetically indistinguishable from those obtained from poultry (meat). Of these ESBL genes, 86% were blaCTX-M-1 and blaTEM-52 genes, which were also the predominant genes in poultry (78%) and retail chicken meat (75%). Of the retail meat samples, 94% contained ESBL-producing isolates of which 39% belonged to E. coli genotypes also present in human samples. These findings are suggestive for transmission of ESBL genes, plasmids and E. coli isolates from poultry to humans, most likely through the food chain.
Identieke resistentiegenen en plasmiden in Escherichia coli van Nederlandse patiënten, pluimvee en kippenvlees
Leverstein-van Hall, M.A. ; Dierikx, C.M. ; Cohen Stuart, J. ; Voets, G.M. ; Munckhof, T.P. Van den; Platteel, T.N. ; Fluit, A.C. ; Sande-Bruinsma, N. Van de; Bonten, M.J.M. ; Mevius, D.J. - \ 2011
Nederlands Tijdschrift voor Geneeskunde 155 (2011). - ISSN 0028-2162
Doel Bepalen welke ‘extended’-spectrum-bètalactamase(ESBL)-genen en plasmiden aanwezig zijn in Escherichia coli in vleeskuikens en vers kippenvlees uit Nederland en tevens in een voor Nederland representatieve collectie van klinische E. coli isolaten van patiënten. Opzet Beschrijvend. Methoden ESBL-producerende E. coli-isolaten waren afkomstig van 98 kipfilets, een ESBL-surveillance studie bij vleeskuikens uit 2006 en 516 humane klinische isolaten uit 31 laboratoria verzameld gedurende een periode van 3 maanden in 2009. De distributie werd vastgesteld van ESBL-genen en plasmiden in E. coli aanwezig in vleeskuikens en vers kippenvlees en de gevonden ESBL-genen werden gedefinieerd als ‘kip-geassocieerd’. In een voor Nederland representatieve steekproef van klinische E. coli-isolaten werd het aandeel van kip-geassocieerde ESBL-genen en plasmiden gekwantificeerd. De isolaten werden geanalyseerd met een ESBL-specifieke microarray, een ESBL-gensequentiebepaling, en 2 plasmide-typeringsmethoden: zogenaamde PCR-gebaseerde replicon-typering en plasmide-multi-locussequentietypering. Resultaten 6 ESBL-genen werden aangemerkt als ‘kip-geassocieerd’: bla CTXM-1, bla CTXM-2, bla SHV-2, bla SHV-12, bla TEM-20, bla TEM-52. Van de humane ESBL-producerende isolaten bevatte 35% een kip-geassocieerd ESBL-gen, en 19% een kip-geassocieerd ESBL-gen gelegen op een IncI1-plasmide, dat genetisch niet te onderscheiden was van plasmides afkomstig van vleeskuikens. In humane isolaten met een kip-geassocieerde ESBL’s waren bla CTXM-1 en bla TEM-52 de meest prevalente ESBL-genen (86%), net als in de isolaten van vleeskuikens (78%) en kipfilets (75%). Van de 98 kipfilets bevatte 94% een ESBL producerende E. coli. Conclusie Deze bevindingen zijn suggestief voor overdracht van ESBL producerende E. coli van pluimvee naar de mens via de voedselketen.
Methicillin-resistant Staphylococcus aureus in horses and horse personnel: An investigation of several outbreaks
Duijkeren, E. van; Moleman, M. ; Oldruitenborgh-Oosterbaan, M.M.S. ; Multem, J. ; Troelstra, A. ; Fluit, A.C. ; Wamel, W.J.B. ; Houwers, D.J. ; Neeling, A.J. de; Wagenaar, J.A. - \ 2010
Veterinary Microbiology 141 (2010)1-2. - ISSN 0378-1135 - p. 96 - 102.
field gel-electrophoresis - animals - mrsa - netherlands - community - st398 - colonization - infections - prevalence - chromosome
At the Veterinary Microbiological Diagnostic Center, the Netherlands, the percentage of methicillin-resistant Staphylococcus aureus (MRSA) isolates found in equine clinical samples increased from 0% in 2002 to 37% in 2008. MRSA of spa-type t064, belonging to MLST ST8 and spa-types t011 and t2123, both belonging to the livestock-associated MLST ST398, predominated. During an outbreak of post-surgical MRSA infections in horses at a veterinary teaching hospital in 2006/2007, MRSA isolates of spa-type t2123 were cultured from 7 horses and 4/61 personnel which indicated zoonotic transmission. After intervention the outbreak stopped. However, another outbreak occurred in 2008, where 17 equine MRSA isolates of spa-type t011 (n = 12), t2123 (n = 4), and t064 (n = 1) were found. This time, 16/170 personnel were positive for MRSA with spa-type t011 (n = 11) and t2123 (n = 5). Personnel in close contact with horses were more often MRSA-positive (15/106) than those without (1/64). Screening of horses upon admission showed that 9.3% were MRSA-positive predominantly with spa-type t011. Weekly cross-sectional sampling of all hospitalized horses for 5 weeks showed that 42% of the horses were MRSA-positive at least once, again predominantly with spa-type t011, which suggests that nosocomial transmission took place. Fifty-three percent of the environmental samples were MRSA-positive, including samples from students' and staff members' rooms, and all were spa-type t011. This indicates that humans contribute to spreading the organism. Culturing of samples employing high-salt pre-enrichment performed better than a comparable method without pre-enrichment. Our results show that nosocomial transmission occurs in equine clinics and suggests that personnel play a role in the transmission.
Rapid detection of TEM, SHV and CTX-M extended-spectrum ß-lactamases in Enterobacteriaceae using ligation-mediated amplification with microarray analysis
Cohen Stuart, J. ; Dierikx, C.M. ; Naiemi, N. Al; Karczmarek, A. ; Hoek, A. ; Vos, P. ; Fluit, A.C. ; Scharringa, J. ; Duim, B. ; Mevius, D.J. ; Leverstein-van Hall, M.A. - \ 2010
Journal of Antimicrobial Chemotherapy 65 (2010)7. - ISSN 0305-7453 - p. 1377 - 1381.
resistance
Objectives Fast and adequate detection of extended-spectrum ß-lactamases (ESBLs) is crucial for infection control measures and the choice of antimicrobial therapy. The aim of this study was to develop and evaluate a novel ESBL assay using ligation-mediated amplification combined with microarray analysis to detect the most prevalent ESBLs in Enterobacteriaceae: TEM, SHV and CTX-M. Methods Analysis of the Lahey database revealed that the vast majority of TEM and SHV ESBLs differ from non-ESBL variants in three amino acid positions. TEM ESBLs have at least one of the following amino acid substitutions: R164S/H/C, G238D/N/S and E104K. In SHV ESBLs, one or more of the following substitutions is observed: D179A/N/G, G238S/A and E240K. Oligonucleotide probes were designed to detect these substitutions, covering 95% of ESBL TEM variants and 77% of ESBL SHV variants. In addition, probes were designed to distinguish between CTX-M groups 1, 2, 9 and 8/25. For evaluation of the assay, 212 Enterobacteriaceae isolates with various ß-lactamases were included (n¿=¿106 ESBL positive). Results The sensitivity of the microarray was 101/106 (95%; 95% CI 89%–98%), and the specificity 100% (95% CI 97%–100%) using molecular characterization of ESBLs by PCR and sequencing as reference. Assay performance time was 8 h for 36 isolates. Conclusions This novel commercially available DNA microarray system may offer an attractive option for rapid and accurate detection of CTX-M, TEM and SHV ESBL genes in Enterobacteriaceae in the clinical laboratory.
Characterization of Dutch Staphylococcus aureus from bovine mastitis using a Multiple Locus Variable Number Tandem Repeat Analysis
Ikawaty, R. ; Brouwer, E.C. ; Jansen, M.D. ; Duijkeren, E. van; Mevius, D.J. ; Verhoef, J. ; Fluit, A.C. - \ 2009
Veterinary Microbiology 136 (2009)3-4. - ISSN 0378-1135 - p. 277 - 284.
field gel-electrophoresis - methicillin-resistant - milk - strains - cows - dna
Current typing methods for Staphylococcus aureus have important drawbacks. We evaluated a Multiple Locus Variable Number Tandem Repeat Analysis (MLVA) scheme with 6 loci which lacks most drawbacks on 85 bovine mastitis isolates from The Netherlands. For each locus the number of repeat units (RU) was calculated. Each combination of repeat units was assigned a MLVA-type (MT). We compared the MLVA typing result with Multi Locus Sequence Typing (MLST), spa-typing and Pulsed-Field Gel Electrophoresis (PFGE). MLVA typing resulted in 18 MTs, although 3 loci could not always be amplified. Spa-typing distinguished 10 spa-types including 3 dominant and 2 new types. PFGE showed 5 dominant profiles with 15 related profiles and 6 unique profiles. MLST showed 4 dominant STs. Some types appeared to be bovine specific. The Simpson's Indices of diversity for PFGE, MLST, spa-typing and MLVA were 0.887, 0.831, 0.69 and 0.781, respectively, indicating that discriminatory power of MLVA was between MLST and spa-typing, whereas PFGE displayed the highest discriminatory power. However, MLVA is fast and cheap when compared to the other methods. The Adjusted Rand index and Wallace's coefficient indicated that MLVA was highly predictive for spa-type, but not vice versa. Analysis of the region neighboring SIRU05 showed a difference in the genetic element bordering the repeats of SIRU05 that explained the negative SIRU05 PCRs. PFGE, MLST, and MLVA are adequate typing methods for bovine-associated S. aureus.
Integrons in Escherichia coli from food-producing animals in The Netherlands
Box, A.T. ; Mevius, D.J. ; Schellen, P. ; Verhoef, J. ; Fluit, A.C. - \ 2005
Microbial Drug Resistance-Mechanisms Epidemiology and Disease 11 (2005)1. - ISSN 1076-6294 - p. 53 - 57.
normal intestinal flora - resistance - swine
The presence and character of class 1 integrons in multidrug-resistant Escherichia coli from slaughter animals and meat was determined by integrase-specific PCR and conserved segment PCR-restriction fragment length polymorphism (RFLP). At least five different class 1 integron types were found and three types were shared between hospitalized patients, humans in the community, meat, and slaughter animals. Common integron types indicate that antibiotic resistance genes are exchanged via the food chain between different reservoirs of both human and animal origin.
Assessment of sustainable development of land and water use : a pilot study in Northwest Overijssel, The Netherlands
Cornelissen, A.M.G. ; Teeuw, J. ; Koops, W.J. ; Fluit, N. van der - \ 2000
In: Livestock Farming Systems, integrating animal science advances into the search for sustainability: Proceedings of the fifth international symposium on livestock farming systems , Posieux, 2000 / Gagnaux, D., Daccord, R., Gibbon, A., Poffet, J.R., Wageningen : Wageningen Pers - p. 127 - 133.
A fluoride-insensitive inorganic pyrophosphatase isolated from Methanothrix soehngenii.
Jetten, M.S.M. ; Fluit, T.J. ; Stams, A.J.M. ; Zehnder, A.J.B. - \ 1992
Archives of Microbiology 157 (1992). - ISSN 0302-8933 - p. 284 - 289.
Merkenonderzoek maaischudders en landbouwcirkelmaaiers = Series test of rotary mowers and rotary mower-conditioners
Fluit, J. - \ 1983
Wageningen : IMAG (Publikatie / Instituut voor Mechanisatie, Arbeid en Gebouwen no. 185) - 63 p.
cilindermaaiers - prestatieniveau - kwaliteit - cirkelmaaiers - keermachines - zwadmaaiers - cylinder mowers - performance - quality - rotary mowers - tedders - windrowers
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