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3. Association between antimicrobial usage in livestock and antimicrobial resistance in Escherichia coli isolates from human urinary tract infections in the Netherlands, 2009–2020.

Author

Meijs, Anouk P, Chanamé-Pinedo, Linda E, Deng, Huifang, Veldman, Kees T, Brouwer, Michael S M, Beld, Maaike J C van den, Dierikx, Cindy M, Sanders, Pim, Wullings, Bart, Greeff, Sabine C de, Duijkeren, Engeline van, Franz, Eelco, Pijnacker, Roan, Mughini-Gras, Lapo, and group, ISIS-AR study

Subjects
ESCHERICHIA coli, URINARY tract infections, DOMESTIC animals, ANIMAL populations, DRUG resistance in microorganisms
Abstract

Background In the last decade, veterinary antimicrobial usage (AMU) and antimicrobial resistance (AMR) among indicator bacteria in livestock have decreased substantially in the Netherlands. The extent to which this decrease has affected AMR levels among human infections remains unclear. Objectives To assess the association between AMU in livestock and AMR in Escherichia coli isolates from human urinary tract infections (UTIs). Methods Data on AMR and AMU between 2009 and 2020 from Dutch national surveillance programmes for humans and livestock were used. Associations between AMU in four major livestock sectors and AMR in humans were assessed for 10 antimicrobial classes and the ESBL resistance profile, using logistic regression analysis. Associations between AMU and AMR in livestock, between AMR in livestock and in humans, and between AMU and AMR in humans were also assessed. Results Statistical significance was reached for 16/31 of the tested associations between AMU in livestock and AMR in human E. coli UTIs. Of the significant associations, 11 were positive (OR 1.01–1.24), whereas 5 were negative (OR 0.96–0.99). All associations between human AMU and AMR in E. coli isolates from UTIs were positive and statistically significant. Weak but significant positive correlations were also observed between livestock AMR and human AMR. Conclusions Although several significant associations between AMU in livestock and AMR in human UTIs caused by E. coli were observed, the associations between AMU and AMR were generally stronger within the human and animal populations. This indicates that potential zoonotic spread of AMR in E. coli causing human UTIs from livestock sources is limited. [ABSTRACT FROM AUTHOR]

Published
2024
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5. A European-wide dataset to uncover adaptive traits of Listeria monocytogenes to diverse ecological niches

Author

Félix, Benjamin, Sevellec, Yann, Palma, Federica, Douarre, Pierre Emmanuel, Felten, Arnaud, Radomski, Nicolas, Mallet, Ludovic, Blanchard, Yannick, Leroux, Aurélie, Soumet, Christophe, Bridier, Arnaud, Piveteau, Pascal, Ascensio, Eliette, Hébraud, Michel, Karpíšková, Renáta, Gelbíčová, Tereza, Torresi, Marina, Pomilio, Francesco, Cammà, Cesare, Di Pasquale, Adriano, Skjerdal, Taran, Pietzka, Ariane, Ruppitsch, Werner, Canelhas, Monica Ricão, Papić, Bojan, Hurtado, Ana, Wullings, Bart, Bulawova, Hana, Castro, Hanna, Lindström, Miia, Korkeala, Hannu, Šteingolde, Žanete, Kramarenko, Toomas, Cabanova, Lenka, Szymczak, Barbara, Gareis, Manfred, Oswaldi, Verena, Marti, Elisabet, Seyfarth, Anne-Mette, Leblanc, Jean-Charles, Guillier, Laurent, and Roussel, Sophie

Published
2022
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7. New insights into the epidemiology of Listeria monocytogenes – A cross-sectoral retrospective genomic analysis in the Netherlands (2010–2020)

Author

Coipan, Claudia E., primary, Friesema, Ingrid H. M., additional, van Hoek, Angela H. A. M., additional, van den Bosch, Tijs, additional, van den Beld, Maaike, additional, Kuiling, Sjoerd, additional, Gras, Lapo Mughini, additional, Bergval, Indra, additional, Bosch, Thijs, additional, Wullings, Bart, additional, van der Voort, Menno, additional, and Franz, Eelco, additional

Published
2023
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8. New insights into the epidemiology of Listeria monocytogenes – A cross-sectoral retrospective genomic analysis in the Netherlands (2010–2020)

Author

Coipan, Claudia E., Friesema, Ingrid H.M., van Hoek, Angela H.A.M., van den Bosch, Tijs, van den Beld, Maaike, Kuiling, Sjoerd, Gras, Lapo Mughini, Bergval, Indra, Bosch, Thijs, Wullings, Bart, van der Voort, Menno, Franz, Eelco, Coipan, Claudia E., Friesema, Ingrid H.M., van Hoek, Angela H.A.M., van den Bosch, Tijs, van den Beld, Maaike, Kuiling, Sjoerd, Gras, Lapo Mughini, Bergval, Indra, Bosch, Thijs, Wullings, Bart, van der Voort, Menno, and Franz, Eelco

Abstract

Introduction: Listeriosis, caused by infection with Listeria monocytogenes (Lm), is a relatively rare but severe disease with one of the highest mortality rates among bacterial foodborne illnesses. A better understanding on the degree of Lm clustering, the temporal distribution of the clusters, and their association with the various food sources is expected to lead to improved source tracing and risk-based sampling. Methods: We investigated the genomic epidemiology of Lm in the Netherlands between 2010 and 2020 by analyzing whole-genome-sequencing (WGS) data of isolates from listerioss patients and food sources from nationwide integrated surveillance and monitoring. WGS data of 756 patient and 770 food/environmental isolates was assessed using core-genome multi-locus sequence typing (cgMLST) with Hamming distance as measure for pairwise distances. Associations of genotype with the epidemiological variables such as patient’s age and gender, and systematic use of specific drugs were tested by multinomial logistic regressions. Genetic differentiation of the Lm within and between food categories was calculated based on allele frequencies at the 1701 cgMLST loci in each food category. Results: We confirmed previous results that some clonal complexes (CCs) are overrepresented among clinical isolates but could not identify any epidemiological risk factors. The main findings of this study include the observation of a very weak attribution of Lm types to food categories and a much better attribution to the producer level. In addition, we identified a high degree of temporal persistence of food, patient and mixed clusters, with more than half of the clusters spanning over more than 1 year and up to 10 years. Discussion: Taken together this would indicate that identifying persistent contamination in food production settings, and producers that process a wide variety of raw food produce, could significantly contribute to lowering the Lm disease burden.

Published
2023

9. Cfr and fexA genes in methicillin-resistant from humans and livestock in the Netherlands

Author

Schouls, Leo M, Veldman, Kees, Brouwer, Michael S M, Dierikx, Cindy, Witteveen, Sandra, van Santen-Verheuvel, Marga, Hendrickx, Antoni P A, Landman, Fabian, Hengeveld, Paul, Wullings, Bart, Rapallini, Michel, Wit, Ben, and van Duijkeren, Engeline

Subjects
Infectious-disease epidemiology, Epidemiology
Abstract

Although the Netherlands is a country with a low endemic level of methicillin-resistant Staphylococcus aureus (MRSA), a national MRSA surveillance has been in place since 1989. In 2003 livestock emerged as a major reservoir of MRSA and currently livestock-associated MRSA (clonal complex CC398) make up 25% of all surveillance isolates. To assess possible transfer of resistant strains or resistance genes, MRSA obtained from humans and animals were characterized in detail.

Published
2022

10. cfr and fexA genes in methicillin-resistant Staphylococcus aureus from humans and livestock in the Netherlands

Author

Schouls, Leo M, Veldman, Kees, Brouwer, Michael S M, Dierikx, Cindy, Witteveen, Sandra, van Santen-Verheuvel, Marga, Hendrickx, Antoni P A, Landman, Fabian, Hengeveld, Paul, Wullings, Bart, Rapallini, Michel, Wit, Ben, van Duijkeren, Engeline, Schouls, Leo M, Veldman, Kees, Brouwer, Michael S M, Dierikx, Cindy, Witteveen, Sandra, van Santen-Verheuvel, Marga, Hendrickx, Antoni P A, Landman, Fabian, Hengeveld, Paul, Wullings, Bart, Rapallini, Michel, Wit, Ben, and van Duijkeren, Engeline

Abstract

BACKGROUND: Although the Netherlands is a country with a low endemic level of methicillin-resistant Staphylococcus aureus (MRSA), a national MRSA surveillance has been in place since 1989. In 2003 livestock emerged as a major reservoir of MRSA and currently livestock-associated MRSA (clonal complex CC398) make up 25% of all surveillance isolates. To assess possible transfer of resistant strains or resistance genes, MRSA obtained from humans and animals were characterized in detail.METHODS: The sequenced genomes of 6327 MRSA surveillance isolates from humans and from 332 CC398 isolates from livestock-related samples were analyzed and resistance genes were identified. Several isolates were subjected to long-read sequencing to reconstruct chromosomes and plasmids.RESULTS: Here we show the presence of the multi-resistance gene cfr in seven CC398 isolates obtained from humans and in one CC398 isolate from a pig-farm dust sample. Cfr induces resistance against five antibiotic classes, which is true for all but two isolates. The isolates are genetically unrelated, and in seven of the isolates cfr are located on distinct plasmids. The fexA gene is found in 3.9% surveillance isolates and in 7.5% of the samples from livestock. There is considerable sequence variation of fexA and geographic origin of the fexA alleles.CONCLUSIONS: The rare cfr and fexA resistance genes are found in MRSA from humans and animals in the Netherlands, but there is no evidence for spread of resistant strains or resistance plasmids. The proportion of cfr-positive MRSA is low, but its presence is worrying and should be closely monitored.

Published
2022

11. A European-wide dataset to uncover adaptive traits of Listeria monocytogenes to diverse ecological niches

Author

Korkealä, Hannu J., Karpíšková, Renáta, Gareis, Manfred, Ruppitsch, W., Hurtado, Ana, Lindström, Miia K., Guillier, Laurent, Cammà, Cesare, Hébraud, Michel, Blanchard, Yannick, Pomilio, Francesco, Gelbíčová, Tereza, Roussel, Sophie, Piveteau, Pascal, Bridier, Arnaud, Pietzka, Ariane Tatjana, Soumet, Christophe, Félix, Benjamin, Torresi, Marina, Radomski, Nicolas, Seyfarth, Anne Mette, Skjerdal, Taran, Wullings, Bart A., Di Pasquale, Adriano, Papić, Bojan, Douarre, Pierre Emmanuel, Felten, Arnaud, Mallet, Ludovic V., Palma, Federica, Szymczak, Barbara, Kramarenko, Toomas, Sévellec, Yann, Šteingolde, Žanete, Leroux, Aurélie, Castro, Hanna, Bulawová, Hana, Oswaldi, Verena, Cabanová, Lenka, Ascensio, Eliette, Canelhas, Monica Ricão, Leblanc, Jean Charles, Marti, Elisabet, Food Hygiene and Environmental Health, Departments of Faculty of Veterinary Medicine, University of Helsinki, Helsinki One Health (HOH), Laboratoire de sécurité des aliments de Maisons-Alfort (LSAl), Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Laboratoire de Ploufragan-Plouzané-Niort [ANSES], Laboratoire de Fougères - ANSES, Optimisation des procédés en Agriculture, Agroalimentaire et Environnement (UR OPAALE), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Agroécologie [Dijon], Université de Bourgogne (UB)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Université Bourgogne Franche-Comté [COMUE] (UBFC)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Microbiologie Environnement Digestif Santé (MEDIS), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Clermont Auvergne (UCA), Veterinary Research Institute [Brno] (VRI), Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise Guiseppe Caporale (IZSAM), Partenaires INRAE, Norwegian Veterinary Institute [Oslo], Austrian Agency for Health and Food Safety (AGES), Swedish Food Agency, University of Ljubljana, Neiker-Instituto Vasco de Investigación y Desarrollo Agrario (SPAIN), Instituto Vasco de Investigación y Desarrollo Agrario [Derio] (NEIKER), Basque Research and Technology Alliance (BRTA), Wageningen Food Safety Research, Wageningen (WFSR), State Veterinary Institute (SVU), Department of Food Hygiene and Environmental Health [Helsinki], Faculty of Veterinary Medicine [Helsinki], Helsingin yliopisto = Helsingfors universitet = University of Helsinki-Helsingin yliopisto = Helsingfors universitet = University of Helsinki, Institute of Food Safety, Animal Health and Environment (BIOR), Veterinary and Food Laboratory, 51006 Tartu, Estonia, Department of Molecular Biology and Epizootiology, Veterinary and Food Institute in Dolny Kubin, State Veterinary and Food Institute, NRL for Honeybee Health, 026 01 Dolny Kubin, Slovakia, West Pomeranian University of Technology Szczecin, Ludwig Maximilian University [Munich] (LMU), Freie Universität Berlin, Agroscope, National Food Institute [Lyngby] (Forside), and European Project: 773830, H2020-SFS-2017-1 ,One Health EJP(2018)

Subjects
ecological niches, 600 Technik, Medizin, angewandte Wissenschaften::630 Landwirtschaft::630 Landwirtschaft und verwandte Bereiche, Moleculaire Biologie & AMR, MESH: Listeria monocytogenes, 413 Veterinary science, Molecular Biology & AMR, génétique, Foodborne Diseases, Bacterial genetics, dataset, Listeriosis, bacteria, bactérie, transmission, Computer Science Applications, CONTAMINATION, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, épidémiologie, epidemiology, Statistics, Probability and Uncertainty, niches écologiques, Information Systems, Statistics and Probability, food-borne pathogen, Library and Information Sciences, écosystème, Education, Micro 3: Knowledge and Expertise, SDG 3 - Good Health and Well-being, Team Bacteriology, données de la recherche, Life Science, Animals, Team Bacteriologie, human, microbiologie, SDG 2 - Zero Hunger, sécurité des aliments, MESH: Foodborne Diseases, Ecosystem, Team Bacteriologie, Moleculaire Biologie & AMR, [SDV.GEN.GPO]Life Sciences [q-bio]/Genetics/Populations and Evolution [q-bio.PE], Team Bacteriology, Molecular Biology & AMR, STRAINS, microbiology, fungi, Bacteriology, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, Listeria monocytogenes, SOIL, MESH: Listeriosis, PORK, genetic, humain, Micro 3: Kennis en Expertise, pathogène alimentaire, CLONES
Abstract

Listeria monocytogenes (Lm) is a ubiquitous bacterium that causes listeriosis, a serious foodborne illness. In the nature-to-human transmission route, Lm can prosper in various ecological niches. Soil and decaying organic matter are its primary reservoirs. Certain clonal complexes (CCs) are over-represented in food production and represent a challenge to food safety. To gain new understanding of Lm adaptation mechanisms in food, the genetic background of strains found in animals and environment should be investigated in comparison to that of food strains. Twenty-one partners, including food, environment, veterinary and public health laboratories, constructed a dataset of 1484 genomes originating from Lm strains collected in 19 European countries. This dataset encompasses a large number of CCs occurring worldwide, covers many diverse habitats and is balanced between ecological compartments and geographic regions. The dataset presented here will contribute to improve our understanding of Lm ecology and should aid in the surveillance of Lm. This dataset provides a basis for the discovery of the genetic traits underlying Lm adaptation to different ecological niches.

Published
2022
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16. Prevalence, risk factors and genetic traits of Salmonella Infantis in Dutch broiler flocks

Author

Mughini-Gras, Lapo, primary, van Hoek, Angela H.A.M., additional, Cuperus, Tryntsje, additional, Dam-Deisz, Cecile, additional, van Overbeek, Wendy, additional, van den Beld, Maaike, additional, Wit, Ben, additional, Rapallini, Michel, additional, Wullings, Bart, additional, Franz, Eelco, additional, van der Giessen, Joke, additional, Dierikx, Cindy, additional, and Opsteegh, Marieke, additional

Published
2021
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17. Exposure to quaternary ammonium compounds selects resistance to ciprofloxacin in listeria monocytogenes

Author

Guérin, Alizée, Bridier, Arnaud, Le Grandois, Patricia, Sévellec, Yann, Palma, Federica, Félix, Benjamin, Roussel, Sophie, Soumet, Christophe, Karpíšková, Renáta, Pomelio, Francesco, Skjerdal, Taran, Ricao, Monica, Bojan, Papič, Oevermann, Anna, Wullings, Bart, Bulawová, Hana, Castro, Hanna, Lindström, Miia, Korkeala, Hannu, Šteingolde, Žanete, Szymczak, Barbara, Gareis, Manfred, and Amar, Corinne

Subjects
Micro 3: Knowledge and Expertise, Team Bacteriologie, Moleculaire Biologie & AMR, Didecyl dimethylammonium chloride, Team Bacteriology, Molecular Biology & AMR, Ciprofloxacin, Biocides, Micro 3: Kennis en Expertise, Antimicrobial resistance, Bacterial adaptation, Benza-lkonium chloride, Listeria monocytogenes
Abstract

In this contribution, the antimicrobial susceptibility toward 11 antibiotics and four biocides of a panel of 205 Listeria monocytogenes (Lm) strains isolated from different ecological niches (i.e., food, animals and natural environment) was evaluated. The impact of exposure to biocides on the antibiotic susceptibilities of Lm was also investigated. Lm strains isolated from food exhib-ited overall a lower susceptibility (higher minimal inhibitory concentrations, MIC) for ammonium quaternary compounds (QACs) and peracetic acid (PAC) than strains isolated from animals and natural environments. Conversely, the ecological origins of Lm strains did not significantly affect their susceptibilities towards antibiotics. Interestingly, repeated exposure to QACs recurrently led to a decrease in susceptibility toward ciprofloxacin (CIP), a fluoroquinolone antibiotic, largely used in human medicine. Moreover, these lower levels of susceptibility to CIP remained stable in most Lm strains even after subcultures without biocide selection pressure, suggesting an adaptation involving modifications at the genetic level. Results underlined the ability of Lm to adapt to biocides, especially QACs, and the potential link between this adaptation and the selection of resistance toward critical antibiotics such as ciprofloxacin. These data support a potential role of the extensive use of QACs from “farm to fork” in the selection of biocide and antibiotic resistance in pathogenic bacteria such as Lm.

Published
2021

18. Prevalence, risk factors and genetic traits of Salmonella Infantis in Dutch broiler flocks

Author

IRAS OH Epidemiology Microbial Agents, dIRAS RA-I&I I&I, Dep Infectieziekten Immunologie, LS Moleculaire Afweer, LS IRAS VPH VV (veterinaire volksgezh.), Mughini-Gras, Lapo, van Hoek, Angela H A M, Cuperus, Tryntsje, Dam-Deisz, Cecile, van Overbeek, Wendy, van den Beld, Maaike, Wit, Ben, Rapallini, Michel, Wullings, Bart, Franz, Eelco, van der Giessen, Joke, Dierikx, Cindy, Opsteegh, Marieke, IRAS OH Epidemiology Microbial Agents, dIRAS RA-I&I I&I, Dep Infectieziekten Immunologie, LS Moleculaire Afweer, LS IRAS VPH VV (veterinaire volksgezh.), Mughini-Gras, Lapo, van Hoek, Angela H A M, Cuperus, Tryntsje, Dam-Deisz, Cecile, van Overbeek, Wendy, van den Beld, Maaike, Wit, Ben, Rapallini, Michel, Wullings, Bart, Franz, Eelco, van der Giessen, Joke, Dierikx, Cindy, and Opsteegh, Marieke

Published
2021

19. Quantitative detection of the free-living amoeba 'Hartmannella vermiformis' in surface water by using real-time PCR

Author

Kuiper, Melanie W., Valster, Rinske M., Wullings, Bart A., Boonstra, Harry, Smidt, Hauke, and van der Kooij, Dick

Subjects
Amoeba -- Environmental aspects, Amoeba -- Research, Ribosomal RNA -- Research, Biological sciences
Abstract

'Hartmannella vermiformis' is a free-living amoeba that is a potential host for 'Legionella pneumophila' in warm water systems and cooling towers. The development of a real-time PCR-based method targeting the 18S rRNA gene for the quantitative detection of this amoeba is described.

Published
2006

21. Intracellular proliferation of Legionella pneumophila in Hartmannella verminformis in aquatic biofilms grown on plasticized polyvinyl chloride

Author

Kuiber, Melanie W., Wullings, Bart A., Akkermans, Antoon D. L., Beumer, Rijkelt R., and Kooij, Dick van der

Subjects
Microbial mats -- Research, Legionella pneumophila -- Growth, Legionella pneumophila -- Research, Company growth, Biological sciences
Abstract

The in vivo growth of Legionella pneumophila in protozoa in aquatic biofilms developing at high concentrations on plasticized polyvinyl chloride in a batch system with autoclaved tap water is discussed. L. pneumophila multiplies in trophozoites of Hartmannella vermiformis are present in aquatic biofilms developing on plastic materials in contact with tap water in a batch test system.

Published
2004

23. Surveillance van Listeria monocytogenes in Nederland, 2018

Author

Friesema, I.H.M., Kuiling, Sjoerd, Heck, M., Wullings, Bart, van der Voort, Menno, Freudenburg-de Graaf, W., van den Ende, A., and Franz, E.

Subjects
Life Science
Abstract

Sinds 2008 is listeriose meldingsplichtig. In 2018 zijn 78 patiënten met listeriose geregistreerd, waaronder 7 zwangere vrouwen (9%). Vier volwassenen zijn ten gevolge van de infectie overleden (6%). De meeste listeriosepatiënten hadden ernstig onderliggende aandoeningen en/of gebruikten immunosuppressiva en/of maagzuurremmers. Een aantal risicoproducten werden in 2018 vaker door patiënten geconsumeerd dan in voorgaande jaren. De meest opvallende stijgers zijn corned beef, gerookte zalm, garnalen en kibbeling/lekkerbek. Whole-genome-sequencing (WGS)-gegevens lieten een aantal clusteringen van patiëntisolaten zien en ook waren een aantal patiëntisolaten geclusterd met voedselisolaten. De meeste clusters bestaan uit patiënten uit verschillende jaren bij wie (vrijwel) identieke stammen zijn aangetoond. Er lijkt dus sprake te zijn van stammen die vanuit persisterende bronnen levensmiddelen besmetten. WGS maakt deze nieuwe inzichten mogelijk en biedt ook nieuwe mogelijkheden om de ziektelast van listeriose verder te verminderen vanwege het grotere vermogen om verbanden te leggen tussen de levensmiddelen en patiënten.Listeria monocytogenes is een bacterie die overal in het milieu voorkomt. De bacterie kan zelfs onder ongunstige omstandigheden zoals droogte en lage temperaturen, overleven en groeien. Infectie bij de mens gebeurt voornamelijk via voedsel dat besmet wordt vanuit de productieomgeving. Het aantal mensen dat listeriose oploopt is niet heel groot, maar de ziektelast is door de ernst van de ziekte hoog. (1, 2) In Nederland bestaat er sinds 2005 een laboratoriumsurveillance voor L. monocytogenes en een aangifteplicht sinds december 2008. Sinds 2017 wordt WGS toegepast als standaard typeringsmethode. Daarnaast worden door de Nederlandse Voedsel en Waren Autoriteit (NVWA) jaarlijks diverse risicovolle voedingsmiddelen op L. monocytogenes onderzocht. In deze rapportage presenteren we de gezamenlijke resultaten van 2018 en vergelijken die met elkaar en ten opzichte van voorgaande jaren.

Published
2019

25. Identification of the bacterial population in manganese removal filters

Author

Bruins, Jantinus H., Petrusevski, Branislav, Slokar, Yness M., Wübbels, Gerhard H., Huysman, Koen, Wullings, Bart A., Joris, Koen, Kruithof, Joop C., Kennedy, Maria D., Bruins, Jantinus H., Petrusevski, Branislav, Slokar, Yness M., Wübbels, Gerhard H., Huysman, Koen, Wullings, Bart A., Joris, Koen, Kruithof, Joop C., and Kennedy, Maria D.

Abstract

The aim of this study was to identify bacteria present in ripened manganese removal filters for drinking water production. The bacterial population was identified with 'next generation' DNA sequencing, and specific bacteria were quantified with quantitative polymerase chain reaction (qPCR) and characterized by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis. The 'next generation' DNA sequencing analysis showed a bacteria population shift from the iron oxidizing species Gallionella spp. in the Fe-filter to manganese and nitrite oxidizing species Pseudomonas spp. and Nitrospira spp., respectively, present in the manganese removal filter. qPCR analysis confirmed the presence of a low concentration of the wellknown Mn2-oxidizing species Ps. putida in the manganese removal filter backwash water. Bacteria of the genus Pseudomonas, isolated from backwash water from a manganese removal filter were cultured and identified with MALDI-TOF MS analysis. Amongst others, P. gessardii, P. grimontii, and P. koreensis were identified. The presence of several manganese oxidizing bacteria species in ripened filter media supports the assumption that a microbial consortium is involved in the oxidation of manganese. Understanding the mechanisms by which manganese coating of filter media commences could endorse the creation of conditions favouring Birnessite formation, and possibly help in reducing typically long ripening periods of manganese removal filters with virgin filter media.

Published
2017

26. Detection of protozoan hosts for Legionella pneumophila in engineered water systems by using a biofilm batch test

Author

Valster, Rinske M., Wullings, Bart A., and van der Kooij, Dick

Subjects
Legionella pneumophila -- Genetic aspects, Legionella pneumophila -- Physiological aspects, Microbial mats -- Physiological aspects, Protozoa -- Genetic aspects, Protozoa -- Physiological aspects, Biological sciences
Abstract

Protozoan hosts are identified for Legionella pneumophila by using a biofilm batch test (BBT). In vitro studies are required to confirm their role as hosts for L. pneumophila and unidentified protozoa are implicated as hosts for uncultured Legiomella spp. grown in BBT flasks.

Published
2010

27. Free-living protozoa in two unchlorinated drinking water supplies, identified by phylogenic analysis of 18S rRNA gene sequences

Author

Valster, Rinske M., Wullings, Bart A., Bakker, Geo, Smidt, Hauke, and van der Kooij, Dick

Subjects
Polymerase chain reaction -- Usage, Protozoa -- Genetic aspects, Protozoa -- Physiological aspects, Restriction fragment length polymorphism analysis -- Usage, Ribosomal RNA -- Research, Water quality bioassay, Biological sciences
Abstract

Free-living protozoa are identified in two unchlorinated groundwater supplies by using cultivation-independent molecular approaches and eukaryotic communities are analyzed by using terminal restriction fragment length polymorphism and clone library analyses of partial 18S rRNA gene fragments and a Hartmannella vermiformis-specific quantitative PCR (qPCR). The studies have shown that the application of qPCR is promising for analyzing the relationship between drinking water quality and the presence of specific protozoa.

Published
2009

28. Occurrence and genetic diversity of uncultured Legionella spp. in drinking water treated at temperatures below 15 degrees Celsius

Author

Wullings, Bart A. and Kooij, Dick van der

Subjects
Legionella -- Physiological aspects, Legionella -- Research, Biological diversity -- Research, Bacteriology -- Cultures and culture media, Bacteriology -- Research, Biological sciences
Abstract

A study was conducted on the drinking water, which was treated at temperature below 15 degrees Celsius, to detect the representatives of the genus Legionella. It was observed that the surface water and ground water treated at temperatures below 15 degrees Celsius showed a large diversity of yet-uncultured Legionellae.

Published
2006

33. Identification ofCasuarina-Frankiastrains by use of polymerase chain reaction (PCR) with arbitrary primers

Author

Sellstedt, Anita, Wullings, Bart, Nyström, Ulrika, and Gustafsson, Petter

Subjects
Identification, Genetics, Frankia, Casuarinaceae, Polymorphism, Molecular Biology, Microbiology, Polymerase chain reaction
Abstract

Free-living N2-fixing Frankia strains isolated from Casuarina sp. were investigated for genomic polymorphism. We used six 10-mer oligonucleotides as single arbitrary primers (AP) for the polymerase chain reaction (PCR) in order to amplify random DNA fragments in the genome of free-living Frankia strains. Agarose-gels of the amplified genomic DNA revealed that two of the six arbitrary primers showed polymorphism in the eight different Frankia genomes. Analysis of the AP-PCR products showed 9 polymorphic bands ranging from 4.1-0.60 kb. We conclude that single arbitrary primers can be used to amplify genomic DNA, and that polymorphism can be detected between the amplification products of the different Frankia genomes.

Published
1992
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39. Identification of Casuarina-Frankia strains by use of polymerase chain reaction (PCR) with arbitrary primers

Author

Sellstedt, Anita, Wullings, Bart, Nyström, Ulrika, and Gustafsson, Petter

Abstract

Free-living N2-fixing Frankia strains isolated from Casuarina sp. were investigated for genomic polymorphism. We used six 10-mer oligonucleotides as single arbitrary primers (AP) for the polymerase chain reaction (PCR) in order to amplify random DNA fragments in the genome of free-living Frankia strains. Agarose-gels of the amplified genomic DNA revealed that two of the six arbitrary primers showed polymorphism in the eight different Frankia genomes. Analysis of the AP-PCR products showed 9 polymorphic bands ranging from 4.1–0.60 kb. We conclude that single arbitrary primers can be used to amplify genomic DNA, and that polymorphism can be detected between the amplification products of the different Frankia genomes.

Published
1992
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40. Reducing antimicrobial use in livestock alone may be not sufficient to reduce antimicrobial resistance among human Campylobacter infections: an ecological study in the Netherlands.

Author

Deng H, Chanamé Pinedo LE, Meijs AP, Sanders P, Veldman KT, Brouwer MSM, Wieke AK, Wullings B, van den Beld MJC, de Greeff SC, Dierikx CM, van Duijkeren E, Franz E, Mughini-Gras L, and Pijnacker R

Subjects
Netherlands epidemiology, Animals, Humans, Cattle, Campylobacter coli drug effects, Campylobacter coli isolation & purification, Chickens microbiology, Poultry microbiology, Cattle Diseases microbiology, Cattle Diseases epidemiology, Cattle Diseases drug therapy, Poultry Diseases microbiology, Poultry Diseases epidemiology, Poultry Diseases drug therapy, Campylobacter Infections veterinary, Campylobacter Infections epidemiology, Campylobacter Infections microbiology, Campylobacter Infections drug therapy, Drug Resistance, Bacterial, Campylobacter jejuni drug effects, Campylobacter jejuni isolation & purification, Anti-Bacterial Agents pharmacology, Anti-Bacterial Agents therapeutic use, Livestock microbiology
Abstract

Reducing antimicrobial use (AMU) in livestock may be one of the keys to limit the emergence of antimicrobial resistance (AMR) in bacterial populations, including zoonotic pathogens. This study assessed the temporal association between AMU in livestock and AMR among Campylobacter isolates from human infections in the Netherlands between 2004 - 2020. Moreover, the associations between AMU and AMR in livestock and between AMR in livestock and AMR in human isolates were assessed. AMU and AMR data per antimicrobial class (tetracyclines, macrolides and fluoroquinolones) for Campylobacter jejuni and Campylobacter coli from poultry, cattle, and human patients were retrieved from national surveillance programs. Associations were assessed using logistic regression and the Spearman correlation test. Overall, there was an increasing trend in AMR among human C. jejuni / coli isolates during the study period, which contrasted with a decreasing trend in livestock AMU. In addition, stable trends in AMR in broilers were observed. No significant associations were observed between AMU and AMR in domestically produced broilers. Moderate to strong positive correlations were found between the yearly prevalence of AMR in broiler and human isolates. Reducing AMU in Dutch livestock alone may therefore not be sufficient to tackle the growing problem of AMR in Campylobacter among human cases in the Netherlands. More insight is needed regarding the population genetics and the evolutionary processes involved in resistance and fitness among Campylobacter.

Published
2024
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41. Aeromonas species from non-chlorinated distribution systems and their competitive planktonic growth in drinking water.

Author

van Bel N, van der Wielen P, Wullings B, van Rijn J, van der Mark E, Ketelaars H, and Hijnen W

Abstract

Aeromonas is included in the Dutch Drinking Water Decree as an indicator for elevated microbial regrowth in non-chlorinated drinking water distribution systems (DWDS). The temporal and spatial diversity of Aeromonas species in ten DWDS and their planktonic growth characteristics for different carbon sources was investigated. Genotyping of the gyr B gene of isolates showed a non-systematic temporal and spatial variable prevalence of seven different Aeromonas species in these DWDS and no correlation with AOC-P17/NOX and Aeromonas concentrations. Pure cultures of these seven species showed a high affinity to low concentrations (μg/L) of individual amino acids and fatty acids, compounds associated with biomass. Growth occurred at 0.5 μg-C/L of an amino acid mixture. Growth of a mixed community of A. rivuli , A. salmonicida , A. sobria and A. veronii in drinking water occurred in pasteurized samples, however, no growth and decay occurred in competition with the autochthonous bacteria (non-pasteurized samples). This community also failed to grow in non-pasteurized distribution samples from a location with clear increase in planktonic Aeromonas concentrations in the transported drinking water. For competitive planktonic growth of Aeromonas an amino acid concentration of ≥5 μg-C/L is required. AOC-P17/NOX concentrations showed that such concentrations are not expected in Dutch drinking water. Therefore, we suspect that competitive planktonic growth is not the major cause of the observed non-compliance with the Aeromonas standard in non-chlorinated DWSD. Importance The occurrence of the bacterial genus Aeromonas in non-chlorinated drinking water in the Netherlands is regarded as an indication for elevated microbial regrowth in the distribution system. Identification of the prevalent species in ten distribution systems by genotyping yielded seven different species, with A. rivuli , A. veronii and A. sobria as the most dominant ones. Planktonic growth experiments of pure cultures confirmed former published affinity of Aeromonas for certain biomass compounds (amino and fatty acids). In competition with the autochthonous microflora, however, planktonic growth was not observed, only after addition of a threshold amino acid concentration of 5 μg-C/L. Based on our results and further observations we deduced that planktonic growth of Aeromonas in the DWDS is not very likely. Benthic growth in loose deposits and planktonic release is a more plausible explanation for the observed planktonic increase of Aeromonas ., (Copyright © 2020 American Society for Microbiology.)

Published
2021
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